TY - JOUR. T1 - Nonmuscle myosin light chain kinase activity modulates radiation-induced lung injury. AU - Wang, Ting. AU - Mathew, Biji. AU - Wu, Xiaomin. AU - Shimizu, Yuka. AU - Rizzo, Alicia N.. AU - Dudek, Steven M.. AU - Weichselbaum, Ralph R.. AU - Jacobson, Jeffrey R.. AU - Hecker, Louise. AU - Garcia, Joe GN. PY - 2016/6/1. Y1 - 2016/6/1. N2 - Radiotherapy as a primary treatment for thoracic malignancies induces deleterious effects, such as acute or subacute radiationinduced lung injury (RILI). Although the molecular etiology of RILI is controversial and likely multifactorial, a potentially important cellular target is the lung endothelial cytoskeleton that regulates paracellular gap formation and the influx of macromolecules and fluid to the alveolar space. Here we investigate the central role of a key endothelial cytoskeletal regulatory protein, the nonmuscle isoform of myosin light chain kinase (nmMLCK), in an established murine RILI model. Our results indicate that thoracic ...
TY - JOUR. T1 - Phosphorylation of smooth muscle myosin light chain kinase by Ca2+/calmodulin-dependent protein kinase II. T2 - Comparative study of the phosphorylation sites. AU - Hashimoto, Yoshiaki. AU - Soderling, Thomas. PY - 1990. Y1 - 1990. N2 - Smooth muscle myosin light chain kinase (MLC-kinase) was rapidly phosphorylated in vitro by the autophosphorylated form of Ca2+/calmodulin-dependent protein kinase II (CaM-kinase II) to a molar stoichiometry of 2.77 ± 0.15 associated with a threefold increase in the concentration of calmodulin (CaM) required for half-maximal activation of MLC-kinase. Binding of CaM to MLC-kinase markedly reduced the phosphorylation stoichiometry to 0.21 ± 0.05 and almost completely inhibited phosphorylation of sites in two peptides (32P-peptides P1 and P2) with reduced phosphorylation of peptide P3. By analogy, cAMP-dependent protein kinase phosphorylated MLC-kinase to a stoichiometry of 3.0 or greater in the absence of CaM with about a threefold decrease in the ...
TY - JOUR. T1 - Definition of the Inhibitory Domain of Smooth Muscle Myosin Light Chain Kinase by Site-Directed Mutagenesis. AU - Ito, Masaaki. AU - Guerriero, Vince. AU - Chen, Xiaomin. AU - Hartshorne, David J.. PY - 1991/4/1. Y1 - 1991/4/1. N2 - Site-directed mutagenesis of smooth muscle myosin light chain kinase was applied to define its autoinhibitory domain. Mutants were all initiated at Leu-447 but contained varying lengths of C-terminal sequence. Those containing the complete C-terminal sequence to Glu-972 possessed kinase activities that were calmodulin-dependent. Removal of the putative inhibitory domain by truncation to Thr-778 resulted in generation of a constitutively active (calmodulin-independent) species. Thus, the inhibitory domain lies to the C-terminal side of Thr-778. Truncation to Lys-793 and to Trp-800 also resulted in constitutively active mutants, although the specific activity of the latter was less than the other mutants. None of the truncated mutants bound calmodulin. ...
TY - JOUR. T1 - Thrombin-mediated Focal Adhesion Plaque Reorganization in Endothelium. T2 - Role of Protein Phosphorylation. AU - Schaphorst, Kane L.. AU - Pavalko, Frederick M.. AU - Patterson, Carolyn E.. AU - Garcia, Joe G.N.. PY - 1997/1/1. Y1 - 1997/1/1. N2 - Endothelial cell (EC) gap formation and barrier function are subject to dual regulation by (1) axial contractile forces, regulated by myosin light chain kinase activity, and (2) tethering forces, represented by cell-cell and cell-substratum adhesions. We examined whether focal adhesion plaque proteins (vinculin and talin) and focal adhesion kinase, p125FAK (FAK), represent target regulatory sites involved in thrombin-mediated EC barrier dysfunction. Histologically, thrombin produced dramatic rearrangement of EC actin, vinculin, and FAK in parallel with the evolution of gap formation and barrier dysfunction. Vinculin and talin were in vitro substrates for phosphorylation by EC PKC, a key effector enzyme involved in thrombin-induced EC ...
Telokin, an abundant gizzard protein, inhibited phosphorylation of regulatory light chain when filamentous myosin was used as the substrate but no inhibition was observed with myosin subfragment 1. At physiological telokin-to-myosin molar ratio (1:1), the inhibition amounted to a 3.5-fold reduction in the initial phosphorylation rate whereas at high molar excess of telokin over myosin, we observed an up to 20-fold decrease in this rate. In agreement with previous observations [Shirinsky, Vorotnikow, Birukov, Nanaev, Collinge, Lukas, Sellers and Watterson (1993) J. Biol. Chem. 268, 16578Ő16583], telokin did not inhibit phosphorylation of the isolated regulatory light chain of myosin and only moderately (35%) inhibited that of heavy meromyosin. To gain a better understanding of the mechanism of this inhibition, we investigated the effects of telokin on the recently described [Babiychuk, Babiychuk and Sobieszek (1995) Biochemistry 34, 6366Ő6372] oligomeric properties of smooth-muscle myosin ...
TY - JOUR. T1 - Regulation of myosin light chain kinase and telokin expression in smooth muscle tissues. AU - Herring, B. Paul. AU - El-Mounayri, Omar. AU - Gallagher, Patricia J.. AU - Yin, Feng. AU - Zhou, Jiliang. PY - 2006/11/24. Y1 - 2006/11/24. N2 - The mylk1 gene is a large gene spanning ∼250 kb and comprising at least 31 exons. The mylk1 gene encodes at least four protein products: two isoforms of the 220-kDa myosin light chain kinase (MLCK), a 130-kDa MLCK, and telokin. Transcripts encoding these products are derived from four independent promoters within the mylk1 gene. The kinases expressed from the mylk1 gene have been extensively characterized and function to regulate the activity of nonmuscle and smooth muscle myosin II. Activation of these myosin motors by MLCK modulates a variety of contractile processes, including smooth muscle contraction, cell adhesion, migration, and proliferation. Dysregulation of these processes contributes to a number of diseases. The noncatalytic gene ...
TY - JOUR. T1 - Differential Regulation of Alternatively Spliced Endothelial Cell Myosin Light Chain Kinase Isoforms by p60Src. AU - Birukov, Konstantin G.. AU - Csortos, Csilla. AU - Marzilli, Lisa. AU - Dudek, Steven. AU - Ma, Shwu Fan. AU - Bresnick, Anne R.. AU - Verin, Alexander D.. AU - Cotter, Robert J.. AU - Garcia, Joe G.N.. PY - 2001/3/16. Y1 - 2001/3/16. N2 - The Ca2+/calmodulin-dependent endothelial cell myosin light chain kinase (MLCK) triggers actomyosin contraction essential for vascular barrier regulation and leukocyte diapedesis. Two high molecular weight MLCK splice variants, EC MLCK-1 and EC MLCK-2 (210-214 kDa), in human endothelium are identical except for a deleted single exon in MLCK-2 encoding a 69-amino acid stretch (amino acids 436-505) that contains potentially important consensus sites for phosphorylation by p60Src kinase (Lazar, V., and Garcia, J. G. (1999) Genomics 57, 256-267). We have now found that both recombinant EC MLCK splice variants exhibit comparable ...
Myosin light chain kinase (MLCK) induces contraction of the perijunctional apical actomyosin ring in response to phosphorylation of the myosin light chain. Abnormal expression of MLCK has been observed in respiratory diseases, pancreatitis, cardiovascular diseases, cancer, and inflammatory bowel disease (IBD). The signaling pathways involved in MLCK activation and triggering of endothelial barrier dysfunction are discussed in this review. The pharmacological effects of regulating MLCK expression by inhibitors such as ML-9, ML-7, microbial products, naturally occurring products, and microRNAs are also discussed. The influence of MLCK in inflammatory diseases starts with endothelial barrier dysfunction. The effectiveness of anti-MLCK treatment may depend on alleviation of that primary pathological mechanism. This review summarizes evidence for the potential benefits of anti-MLCK agents in the treatment of inflammatory disease and the importance of avoiding treatment-related side effects, as MLCK is widely
TY - JOUR. T1 - Different phosphorylated forms of myosin in contracting tracheal smooth muscle. AU - Persechini, A.. AU - Kamm, K. E.. AU - Stull, J. T.. PY - 1986. Y1 - 1986. N2 - Calmodulin-dependent myosin light chain kinase phosphorylates two light chain subunits on each myosin molecule. We have developed a method for measuring nonphosphorylated, monophosphorylated, and diphosphorylated forms of myosin in smooth muscle. Four protein bands were separated in tissue extracts by nondenaturing polyacrylamide gel electrophoresis in the presence of pyrophosphate. Immunoblots demonstrated that three forms (designated M, MP, and MP2) reacted with rabbit antisera prepared against the purified phosphorylated light chain (P-light chain) from bovine tracheal smooth muscle. Evidence was obtained that M, MP, and MP2 represented nonphosphorylated, monophosphorylated, and diphosphorylated myosin, respectively, and that the other protein band was probably filamin. The formation of different phosphorylated ...
A major focus of research in my laboratory deals with understanding the role and regulation of actin-myosin interactions in endothelial cell (EC) inflammation, permeability, and apoptosis, especially in the settings of acute and chronic lung injury. We study pathways of EC inflammation, barrier dysfunction and apoptosis on the molecular, cellular, and animal level. We have uncovered novel and previously unrecognized role of actin cytoskeleton and non-muscle myosin light chain kinase (nmMLCK) in the regulation of cytoplasmic trafficking of NF-k? for its nuclear import to cause EC inflammation associate with intravascular coagulation and sepsis. More recently we have been involved in the identification of endoplasmic reticulum (ER) stress as an important determinant of EC inflammation and permeability associated with acute lung injury (ALI). In our most recent work, we have focused on understanding the role of nmMLCK in the pathogenesis of pulmonary emphysema caused by cigarette smoke (CS). The ...
Cardiac-specific myosin light chain kinase (cMLCK) is the kinase predominantly responsible for the maintenance of the basal level of phosphorylation of cardiac myosin light chain 2 (MLC2), which it phosphorylates at Ser-15. This phosphorylation repels the myosin heads from the thick myosin filament and moves them toward the thin actin filament. Unlike smooth muscle cells, MLC2 phosphorylation in striated muscle cells appears to be a positive modulator of Ca2+ sensitivity that shifts the Ca2+-force relationship toward the left and increases the maximal force response and thus does not initiate muscle contraction. Recent studies have revealed an increasing number of details of the biochemical, physiological, and pathophysiological characteristics of cMLCK. The combination of recent technological advances and the discovery of a novel class of biologically active nonstandard peptides will hopefully translate into the development of drugs for the treatment of heart diseases. (Circ J 2013; 77: ...
The Ca(2+)-dependent kinase myosin light chain kinase (MLCK) is the activator of smooth muscle contraction. In addition, it has been reported to be involved in Ca(2+) channel regulation in cultured cells, and we previously showed that the MLCK inhibitor ML-7 decreases arginine vasopressin (AVP)-induced Ca(2+) influx in rat aorta. This study was designed to investigate whether MLCK is involved in Ca(2+) regulation in resistance artery smooth muscle cell, which plays a major role in the control of blood pressure. As ML compounds were shown to have off-target effects, MLCK was downregulated by transfection with a small interfering RNA targeting MLCK (MLCK-siRNA) in rat small resistance mesenteric artery (RMA) and in the rat embryonic aortic cell line A7r5. Noradrenaline-induced contraction and Ca(2+) signal were significantly depressed in MLCK-siRNA compared to scramble-siRNA-transfected RMA. Contraction and Ca(2+) signal induced by high KCl and voltage-activated Ca(2+) current were also ...
Circular RNA Myosin Light Chain Kinase (MYLK) Promotes Prostate Cancer Progression through Modulating Mir-29a Expression - Article statistics #908009
TY - JOUR. T1 - Adherent neutrophils activate endothelial myosin light chain kinase. T2 - Role in transendothelial migration. AU - Garcia, Joe G.N.. AU - Verin, Alexander D.. AU - Herenyiova, Maria. AU - English, Denis. PY - 1998/5/1. Y1 - 1998/5/1. N2 - Increased vascular endothelial cell (EC) permeability and neutrophilic leukocyte (PMN) diapedesis through paracellular gaps are cardinal features of acute inflammation. Activation of the EC contractile apparatus is necessary and sufficient to increase vascular permeability in specific models of EC barrier dysfunction. However, it is unknown whether EC contraction with subsequent paracellular gap formation is required for PMN transendothelial migration in response to chemotactic factors. To test this possibility, we assessed migration of human PMNs across confluent bovine pulmonary arterial EC monolayers. Transendothelial PMN migration in the absence of a chemotactic gradient was minimal, whereas abluminal addition of leukotriene B4 (LTB4; 5 μM) ...
Reactome is pathway database which provides intuitive bioinformatics tools for the visualisation, interpretation and analysis of pathway knowledge.
Myosin regulatory light polypeptide 9 is a protein that in humans is encoded by the MYL9 gene. Myosin, a structural component of muscle, consists of two heavy chains and four light chains. The protein encoded by this gene is a myosin light chain that may regulate muscle contraction by modulating the ATPase activity of myosin heads. The encoded protein binds calcium and is activated by myosin light chain kinase. Two transcript variants encoding different isoforms have been found for this gene. GRCh38: Ensembl release 89: ENSG00000101335 - Ensembl, May 2017 GRCm38: Ensembl release 89: ENSMUSG00000067818 - Ensembl, May 2017 "Human PubMed Reference:". "Mouse PubMed Reference:". Kumar CC, Mohan SR, Zavodny PJ, Narula SK, Leibowitz PJ (May 1989). "Characterization and differential expression of human vascular smooth muscle myosin light chain 2 isoform in nonmuscle cells". Biochemistry. 28 (9): 4027-35. doi:10.1021/bi00435a059. PMID 2526655. "Entrez Gene: myosin". Stelzl U, Worm U, Lalowski M, et al. ...
MYLK2 - MYLK2 (untagged)-Kinase deficient mutant (K314M) of Human myosin light chain kinase 2 (MYLK2) available for purchase from OriGene - Your Gene Company.
Complete information for MYLK4 gene (Protein Coding), Myosin Light Chain Kinase Family Member 4, including: function, proteins, disorders, pathways, orthologs, and expression. GeneCards - The Human Gene Compendium
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CBP affinity tag is a useful tag in protein study which can be added to the N or C terminus of proteins of interest through DNA recombinant technology. The tag is derived from muscle myosin light-chain kinase. The tag co,,
Expresses constitutively active chicken myosin light chain kinase under UAS control, M. V. Identity of balancer is a guess; it may be TM3, P{ftz/lacC}SC1, Sb[1] ry[RK]; homozygotes present, K.C ...
Intrahepatic cholestasis represents 20%-40%of drug-induced injuries from which a large proportion remains unpredictable. We aimed to investigate mechanisms underlying drug-induced cholestasis and improve its early detection using human HepaRG cells and a set of 12 cholestatic drugs and six noncholestatic drugs. In this study, we analyzed bile canaliculi dynamics, Rho kinase (ROCK)/myosin light chain kinase (MLCK) pathway implication, efflux inhibition of taurocholate [a predominant bile salt export pump (BSEP) substrate], and expression of the major canalicular and basolateral bile acid transporters. We demonstrated that 12 cholestatic drugs classified on the basis of reported clinical findings caused disturbances of both bile canaliculi dynamics, characterized by either dilatation or constriction, and alteration of the ROCK/MLCK signaling pathway, whereas noncholestatic compounds, by contrast, had no effect. Cotreatment with ROCK inhibitor Y-27632 [4-(1-aminoethyl)-N-(4-pyridyl) cyclohexanecarboxamide
Objective: This study investigated signaling pathways that may contribute to the potent positive inotropic effect of human urotensin-II (hU-II) in human isolated right atrial trabeculae obtained from patients with coronary artery disease. Methods: Trabeculae were set up in tissue baths and stimulated to contract at 1 Hz. Tissues were incubated with 20 nM hU-II with or without phorbol 12-myristate 13-acetate (PMA, 10 μM) to desensitize PKC, the PKC inhibitor chelerythrine (10 μM), 10 μM 4α-phorbol that does not desensitize PKC, the myosin light chain kinase inhibitor wortmannin (50 nM, 10 μM), or the Rho kinase inhibitor Y-27632 (0.1-10 μM). Activated RhoA was determined by affinity immunoprecipitation, and phosphorylation of signaling proteins was determined by SDS-PAGE. Results: hU-II caused a potent positive inotropic response in atrial trabeculae, and this was concomitant with increased phosphorylation of regulatory myosin light chain (MLC-2, 1.8±0.4-fold, P less than 0.05, n=6) and PKCα/βII
The vertebrate genetic locus, coding for a Ca2+/calmodulin-dependent enzyme myosin light chain kinase (MLCK), the key regulator of smooth muscle contraction and cell motility, reveals a complex organization. Two MLCK isoforms are encoded by the MLCK genetic locus. Recently identified M(r) 210 kDa MLCK contains a sequence of smooth muscle/non-muscle M(r) 108 kDa MLCK and has an additional N-terminal sequence (Watterson et al., 1995. FEBS Lett. 373 : 217). A gene for an independently expressed non-kinase product KRP (telokin) is located within the MLCK gene (Collinge et al., 1992. Mol. Cell. Biol. 12 : 2359). KRP binds to and regulates the structure of myosin filaments (Shirinsky et al., 1993. J. Biol. Chem. 268 : 16578). Here we compared biochemical properties of MLCK-210 and MLCK-108 and studied intracellular localization of MLCK-210. MLCK-210 was isolated from extract of chicken aorta by immunoprecipitation using specific antibody and biochemically analysed in vitro. MLCK-210 phosphorylated myosin
BACKGROUND: Intestinal epithelial barrier dysfunction, which involves myosin light chain kinase (MLCK) activation, contributes to the occurrence and progression of inflammation in inflammatory bowel disease (IBD). Wogonoside helps maintain intestinal homeostasis in mice with dextran sulfate sodium (DSS)-induced colitis, but it is unclear whether it modulates intestinal barrier function. PURPOSE: Here, we demonstrate that wogonoside protects against intestinal barrier dysfunction in colitis via the MLCK/pMLC2 pathway both in vivo and in vitro. METHODS: Caco-2 cell monolayers treated with the proinflammatory cytokine TNF-α showed barrier dysfunction and were assessed in the absence and presence of wogonoside for various physiological, morphological, and biochemical parameters. Colitis was induced by 3% DSS in mice, which were used as an animal model to explore the pharmacodynamics of wogonoside. We detected MLCK/pMLC2 pathway proteins via western blot analysis, assessed the cytokines IL-13 and ...
6. Abstract Ethanol is a common factor in traumatic injury, including burn injury. Previous studies from our laboratory indicate that ethanol increases both pul...
Rabbit recombinant monoclonal Myosin light chain kinase antibody [EP1458Y] validated for WB, IHC, Flow Cyt, ICC/IF and tested in Human, Mouse and Rat…
Mounting evidence shows that the urokinase plasminogen activator (uPA) and its receptor (uPAR) play a central role in tumor progression. 120 h p.i. In vivo uPA specificity of 89Zr-Df-ATN-291 was confirmed by successful pharmacological blocking of tumor uptake with ATN-291 in U87MG tumors. Akap7 Although the detailed mechanisms behind [19], and more recently, an 111In-labeled antibody adopted for prostate cancer imaging with remarkable tumor accumulation [20]. Our goal was to investigate a novel probe for effective targeting and imaging of the uPA/uPAR system in cancer with excellent targeting specificity and image contrast. To achieve this goal, 89Zr-labeled Bortezomib ATN-291 (i.e., 89Zr-Df-ATN-291; Df is abbreviated for deferoxamine) was used as an immunoPET probe. 89Zr (t1/2 = 78.4 h) was selected as the radiolabel in this study to provide a longitudinal evaluation on the interaction between ATN-291 and different tumor types [21]. To accomplish this goal, various studies were carried out to ...
Expression of MYLK (MLCK, MLCK1, MYLK1, smMLCK) in cancer tissue. The cancer tissue page shows antibody staining of the protein in 20 different cancers.
Expression of MYLK (MLCK, MLCK1, MYLK1, smMLCK) in prostate tissue. Antibody staining with HPA031677 and CAB020789 in immunohistochemistry.
This golden turmeric mylk latte, also known as golden mylk, is a healing, cozy, dairy-free drink with anti-inflammatory benefits.
This golden turmeric mylk latte, also known as golden mylk, is a healing, cozy, dairy-free drink with anti-inflammatory benefits.
Complete information for MYLKP1 gene (Pseudogene), Myosin Light Chain Kinase Pseudogene 1, including: function, proteins, disorders, pathways, orthologs, and expression. GeneCards - The Human Gene Compendium
Gene target information for Mylk2 - myosin, light polypeptide kinase 2, skeletal muscle (house mouse). Find diseases associated with this biological target and compounds tested against it in bioassay experiments.
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Inhibitor studies have indicated that MLCK activation, as assessed by MLC phosphorylation, is necessary for TJ regulation secondary to diverse physiological and pathophysiological stimuli (Hopkins et al., 2003; Scott et al., 2002; Turner et al., 2000; Turner et al., 1997; Yuhan et al., 1997; Zolotarevsky et al., 2002). However, the nonspecific nature of pharmacologic agents, the complexity of the stimuli used, and the absence of appropriate experimental models has made it impossible to determine if MLC phosphorylation alone is sufficient to trigger acute downstream regulation of assembled TJs. To address this problem, we developed a model of inducible tMLCK expression in fully differentiated epithelial monolayers, thereby overcoming limitations of previous studies (Gandhi et al., 1997; Hecht et al., 1996). Our data show that induction of tMLCK expression in mature monolayers causes MLC phosphorylation and is sufficient to cause increases in TJ permeability. These changes in TJ function are ...
TY - JOUR. T1 - Fluorescence analysis of calmodulin mutants containing tryptophan. T2 - Conformational changes induced by calmodulin-binding peptides from myosin light chain kinase and protein kinase II. AU - Prendergast, Franklyn G.. PY - 1991. Y1 - 1991. N2 - Peptide-induced conformational changes in five isofunctional mutants of calmodulin (CaM), each bearing a single tryptophan residue either at the seventh position of each of the four calcium-binding loops (i.e., amino acids 26, 62, 99, and 135) or in the central helix (amino acid 81) were studied by using fluorescence spectroscopy. The peptides RS20F and RS20CK correspond to CaM-binding amino acid sequence segments of either nonmuscle myosin light chain kinase (nmMLCK) or calmodulin-dependent protein kinase II (CaMPK-II), respectively. Both steady-state and time-resolved fluorescence data were collected from the various peptide-CaM complexes. Steady-state fluorescence intensity measurements indicated that, in the presence of an excess of ...
Title: Regulatory Light Chains of Striated Muscle Myosin. Structure, Function and Malfunction. VOLUME: 3 ISSUE: 2. Author(s):Danuta Szczesna-Cordary. Affiliation:Department of Molecular&Cellular Pharmacology, University of Miami School of MedicineRosenstiel Medical Sciences Building R-189, Room 6113, USA. Keywords:regulatory light chains of myosin (rlc), Phosphorylation, skinned fibers, familial hypertrophic, cardiomyopathy (fhc) mutations.. Abstract: Striated (skeletal and cardiac) muscle is activated by the binding of Ca2+ to troponin C and is regulated by the thin filament proteins, tropomyosin and troponin. Unlike in molluscan or smooth muscles, the myosin regulatory light chains (RLC) of striated muscles do not play a major regulatory role and their function is still not well understood. The N-terminal domain of RLC contains a Ca2+-Mg2+-binding site and, analogous to that of smooth muscle myosin, also contains a phosphorylation site. During muscle contraction, the increase in Ca2+ ...
Contraction of smooth muscle cells, such as those that line blood vessels, is regulated through phosphorylation of the myosin light chain. Phosphorylation by myosin light-chain kinase (and consequent increased contraction) is opposed by dephosphorylation (and consequent relaxation) catalyzed by the myosin light-chain phosphatase (PP1M). Some agents that control blood pressure act by modulating this regulatory system. Nitric oxide, a vasodilator, causes increased intracellular production of cyclic guanosine monophosphate (cGMP), which in turn activates cGMP-dependent protein kinase (cGKIα). Surks et al. provide evidence that cGKIα localizes as part of a multienzyme complex at the contractile apparatus through direct interaction with the myosin-binding subunit (MBS) of PP1M. This association was required for cGMP-dependent activation of PP1M and dephosphorylation of myosin light chain. The CGKIα enzyme adds yet another component to the cluster of regulatory components bound to the MBS, which ...
The study by Sato et al9 raises important questions of how the RhoA-Rho-kinase pathway is activated after the insult of hemorrhage, how MLCP activity is regulated in general, and whether these mechanisms are altered in various pathological states. MLCP is affected not only by various signaling cascades that might alter the state of phosphorylation of MBS, but there are also multiple sites for regulation by protein factors that could modify the interactions among the subunit domains involved in MLCP targeting and catalysis. There is evidence for modulation of MLCP activity by either activator or repressor proteins, whose activity is modified by phosphorylation. A potential activator protein is telokin, which comprises an independently expressed C-terminal domain of MLCK.12 13 It is not clear whether telokin acts by modifying the activity of MLCP directly or indirectly by interactions with MLC. Whatever the case, its activity as a promoter of MLCP activity is amplified when telokin is ...
TY - JOUR. T1 - Dynamic regulation of vascular myosin light chain (MYL9) with injury and aging. AU - Shehadeh, Lina A.. AU - Webster, Keith A.. AU - Hare, Joshua M.. AU - Vazquez-Padron, Roberto I.. PY - 2011/10/7. Y1 - 2011/10/7. N2 - Background: Aging-associated changes in the cardiovascular system increase the risk for disease development and lead to profound alterations in vascular reactivity and stiffness. Elucidating the molecular response of arteries to injury and age will help understand the exaggerated remodeling of aging vessels. Methodology/Principal Findings: We studied the gene expression profile in a model of mechanical vascular injury in the iliac artery of aging (22 months old) and young rats (4 months old). We investigated aging-related variations in gene expression at 30 min, 3 d and 7 d post injury. We found that the Myosin Light Chain gene (MYL9) was the only gene differentially expressed in the aged versus young injured arteries at all time points studied, peaking at day 3 ...
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Fasudil, Monohydrochloride is a cell-permeable Ca²⁺ antagonist and vasodilator. Inhibits protein kinase A (Ki = 1.6 µM), protein kinase G (Ki = 1.6 µM), myosin light chain kinase (Ki = 36 µM) and Rho kinase (ROCK; IC₅₀ = 10.7 µM).
After obtaining a BS in Biochemistry at the University of Wisconsin-Madison, Chew went to St. Louis University to pursue his PhD, where he worked to understand the role of myosin II regulation in endothelial cells. Realizing that conventional biochemical methods could not address the spatial and temporal regulation of signaling pathways inside the cell, Chew embarked on his postdoctoral research in the laboratory of Rex Chisholm by developing fluorescent biosensor to simultaneously monitor the enzymatic activity and localization pattern of myosin light chain kinase in vivo.
Food allergies, celiac disease, inflammatory bowel disease (IBD), diarrhea and other gastrointestinal diseases have something in common: all have been linked to epithelial barrier loss. The gut epithelial barrier-that critical lining of cells in the gut that must allow nutrients into the body while keeping food-borne microbes out-can be compromised during intestinal inflammation and cause disease. While many of the molecular mechanisms that trigger gastrointestinal diseases remain a mystery, previous research has found that one enzyme, known as myosin light chain kinase (MLCK), plays a critical role. However, MLCK is also essential for critical functions in gut epithelia and other cell types. This makes direct inhibition of MLCK impossible, as it would result in many toxic and systemic side effects. A team led by investigators from Brigham and Womens Hospital has now developed an alternative approach. In a study published in Nature Medicine, the researchers report new evidence suggesting that ...
Buy anti-PRKDC antibody, Rabbit Myosin Light Chain 2 Polyclonal Antibody-NP_001075109.1 (MBS857334) product datasheet at MyBioSource, Primary Antibodies. Application: Immunohistochemistry (IHC)
Abstract: Myosin heavy chain kinase (MHCK) A phosphorylates mapped sites at the C-terminal tail of Dictyostelium myosin II heavy chain, driving disassembly of myosin filaments both in vitro and in vivo. MHCK A is organized into three functional domains that include an N-terminal coiled-coil region, a central kinase catalytic domain unrelated to conventional protein kinases, and a WD repeat domain at the C terminus. MHCK B is a homologue of MHCK A that possesses structurally related catalytic and WD repeat domains. In the current study, we explored the role of the WD repeat domains in defining the activities of both MHCK A and MHCK B using recombinant bacterially expressed truncations of these kinases either with or without their WD repeat domains. We demonstrate that substrate targeting is a conserved function of the WD repeat domains of both MHCK A and MHCK B and that this targeting is specific forDictyostelium myosin II filaments. We also show that the mechanism of targeting involves direct ...
Myosin light chain kinase (MLCK)-dependent phosphorylation of the regulatory light chain (RLC) of cardiac myosin is known to play a beneficial role in heart disease, but the idea of a phosphorylation-mediated reversal of a hypertrophic cardiomyopathy (HCM) phenotype is novel. Our previous studies on transgenic (Tg) HCM-RLC mice revealed that the D166V (Aspartate166 →Valine) mutation-induced changes in heart morphology and function coincided with largely reduced RLC phosphorylation in situ. In this paper, we hypothesized that the introduction of a constitutively phosphorylated Serine15 (S15D) into the hearts of D166V mice would prevent the development of a deleterious HCM phenotype. In support of this notion, MLCK-induced phosphorylation of D166V-mutated hearts was found to rescue some of their abnormal contractile properties. Tg-S15D-D166V mice were generated with the human cardiac RLC-S15D-D166V construct substituted for mouse cardiac RLC and were subjected to functional, structural, and ...
Although cytoskeletal proteins such as myosin II are implicated in the control of insulin secretion, their precise role is poorly understood. In other secretory cells, myosin II is predominantly regulated via the phosphorylation of the regulatory light chains (RLC). The current study was aimed at investigating RLC phosphorylation in beta-cells. In both the insulin-secreting cell line RINm5F and rat pancreatic islets, the RLC was basally phosphorylated on the myosin light chain kinase sites (Ser19/Thr18). Phosphorylation at these sites was not consistently increased by either metabolic stimuli (glyceraldehyde/glucose) or the depolarizing agent KCl. The RLC sites phosphorylated by protein kinase C (PKC) (Ser1/Ser2) were unphosphorylated in the basal state, not affected by nutrients or KCl, and only slightly increased by the PKC activator phorbol 12-myristate 13-acetate (PMA). Like the other insulin secretagogues, however, PMA did promote serine phosphorylation of the myosin heavy chain (MHC) in RINm5F
During cell migration, myosin II modulates adhesion, cell protrusion and actin organization at the leading edge. We show that an F-actin- and membrane-associated scaffolding protein, called supervillin (SV, p205), binds directly to the subfragment 2 domains of nonmuscle myosin IIA and myosin IIB and to the N-terminus of the long form of myosin light chain kinase (L-MLCK). SV inhibits cell spreading via an MLCK- and myosin II-dependent mechanism. Overexpression of SV reduces the rate of cell spreading, and RNAi-mediated knockdown of endogenous SV increases it. Endogenous and EGFP-tagged SV colocalize with, and enhance the formation of, cortical bundles of F-actin and activated myosin II during early cell spreading. The effects of SV are reversed by inhibition of myosin heavy chain (MHC) ATPase (blebbistatin), MLCK (ML-7) or MEK (U0126), but not by inhibiting Rho-kinase with Y-27632. Flag-tagged L-MLCK co-localizes in cortical bundles with EGFP-SV, and kinase-dead L-MLCK disorganizes these bundles. The L