Mechanism mutagenesis site directed. Medical search. Frequent questions

Significant insight into their catalytic mechanisms has been obtained by relating site-directed mutagenesis studies to kinetic activity assays. (iucr.org)

Variants produced by substitution of Cys(972) and Cys(353) by Ser using site-directed mutagenesis have distinctly lower specific activities, supporting the crucial role of these highly conserved redox-sensitive amino acid residues for P-protein activity. (nih.gov)
Structure-guided mutagenesis reveals key RNA contact residues that are critical for RocC/RocR to repress the uptake of environmental DNA in L. pneumophila . (nature.com)
Surface-exposed residues, Arg24, Lys27, and Arg31 of the beta-sheet, have been identified from mutagenesis studies as functionally important for blocking the Shaker K+ channel. (rcsb.org)
He has also defined, by site-directed mutagenesis, essential amino acid residues concerned in phosphate transport and transmembrane signalling in the pho regulon in Escherichia coli. (science.org.au)
The association of potential effector proteins with the VEGFRs were assessed in both yeast and mammalian cells, and the identity of the interacting residues was probed using site-directed mutagenesis and peptide competition experiments. (figshare.com)
By site-directed mutagenesis, we have identified several amino acid residues on cten and DLC-1 that are essential for this interaction. (rupress.org)

Also called site-specific mutagenesis or oligonucleotide-directed mutagenesis, it is used for investigating the structure and biological activity of DNA, RNA, and protein molecules, and for protein engineering. (wikipedia.org)
The working group prepared a list of seven new plant breeding techniques: zinc finger nuclease (ZFN) technology, oligonucleotide-directed mutagenesis (ODM), cisgenesis and intragenesis, grafting on GM-rootstock, RNA-dependent DNAmethylation, agro-infiltration "sensu stricto," and reverse breeding. (frontiersin.org)

The experiments showed that in the inactive form, the two ends of the protein form a sheet that closes a key site on the protein and prevents it from changing into its active form. (elifesciences.org)
His recent site-directed mutagenesis experiments on the mechanism of oxidative phosphorylation have solved the long-standing problem of the nature of the proton channel of the ATP synthase. (science.org.au)
Site directed mutagenesis experiments showed that reducing the positive charge of this surface-exposed region of the E protein DII correlated with improved Ae. (cdc.gov)

PLP and the substrate glycine bind in the active site of this reduced enzyme and appear to cause further conformational changes involving a flexible surface loop. (nih.gov)
For advances in protein structure and enzyme mechanisms by use of affinity labeling and site-directed mutagenesis. (ornl.gov)
Here, a systematic approach to investigate the effect of an active-site-residue mutation on a model enzyme, human carbonic anhydrase II (CA II), is described. (iucr.org)
Enzyme active sites provide highly optimized microenvironments for their specific substrates by providing reactive groups such as nucleophiles or acids/bases that stabilize the transition state. (iucr.org)
The second aim consists of establishing an innovative and reproducible coupling mechanism between a specific antibody and the selected enzyme, which are used together to report the virus antigen. (fhnw.ch)
To achieve these goals we use site-directed mutagenesis of the enzymes, enzyme expression and purification and various enzyme activity tests. (fhnw.ch)
Alternative sites for cell density regulation of the enzyme could rely on its sensitivity to sulfhydryl groups. (embl.de)
The studies include, enzyme kinetic studies, mechanism, expression in bacteria, mutant enzymes by site-directed mutagenesis of the gene, partial sequencing of the genomic DNA (upstream region), gene replacement in fibroblasts and studies towards preparing a DHPR 'knockout mouse' as a model for the disease for gene replacement. (edu.au)
In order to investigate the interchain contacts involved in the cooperative interactions between the catalytic sites, a series of modified forms of the enzyme was prepared by site directed mutagenesis. (europa.eu)
The results obtained show that the R1-C4 interaction is essential for the establishment of the enzyme conformation having low affinity for aspartate (T state), and consequently for the existence of cooperactivity between the catalytic sites. (europa.eu)
Some sulfur- Here we report the discovery and characterization of a pre- oxidizing bacteria (SOB) known to play an important role in viously undescribed copper center that forms the active site of global geochemical cycles can utilize thiocyanate as the sole a copper-containing enzyme thiocyanate dehydrogenase (sug- source of energy and nitrogen . (docslib.org)
Over 20 copper-containing enzymes that the active site of a not previously described enzyme, thiocya- catalyze a wide-ranging group of chemical reactions have been nate dehydrogenase. (docslib.org)

The results establish that C his 134 is indeed the residue interacting with the carbonyl group of CP but that it is not directly involved in the catalytic mechanism. (europa.eu)

Early attempts at mutagenesis using radiation or chemical mutagens were non-site-specific, generating random mutations. (wikipedia.org)

Photoaffinity labeling with a fluorescent nonpolar peptide caused the covalent attachment of the peptide to a single site on the Tsp protein. (embl.de)
Site-directed mutagenesis of a surface residue at the peptide binding site of the PDZ domain, valine 229, to Glu or Gln resulted in an increase in the K(M) value but had no effect on the k(cat) value. (embl.de)
1995), the molecular mechanism and the peptide group, suggesting that urea-peptide inter- whereby urea unfolds proteins has not been established. (lu.se)

Routine biochemistry studies of catalytic mechanisms, for instance, routine studies using tools such as site-directed mutagenesis, are not typically suitable. (rsc.org)

The methodology will include site-directed mutagenesis, covalent protein modifications, patch- clamp measurements, thermodynamic and kinetic analysis of dose-response curves and rates of transitions, videomicroscopy, and molecular modeling. (usda.gov)

Various techniques in molecular biology (Biochemical characterization of enzymes, protein purification, SDS-PAGE, Western blotting, construction of appropriate vectors and cloning, site directed mutagenesis etc. (uni-goettingen.de)
L-Methionine γ-lyase from Pseudomonas putida has a conserved tyrosine residue (Tyr114) in the active site as in all known sequences of γ-family pyridoxal 5′-phosphate dependent enzymes. (elsevierpure.com)

Systematic deletion mutagenesis of Tsp localized the binding site to amino acids 206-307, a region that completely encompasses the putative PDZ domain (217-301). (embl.de)

02/2010 - 04/2011: Site-directed mutagenesis of alpha-amylase and characterization of biochemical (kinetic and activation) parameters of mutants and wild type. (uni-goettingen.de)
Transposon mutagenesis has been proven to be a powerful tool to identify genes and networks which are involved in survival and fitness under a given condition by simultaneously assaying the fitness of millions of mutants, thereby relating genotype to phenotype and contributing to an understanding of bacterial cell biology. (biorxiv.org)
Studies of these mutants led to new insight into the pump's mechanism and provided an improved understanding of the genetic basis for human skeletal muscle conditions such as Brody myopathy and malignant hyperthermia. (royalsociety.org)

Site directed mutagenesis was used to check the catalytic role of residue C his 134, which is in a position to interact with the carbonyl group of CP, and was a candidate for bearing a group whose deprotonation is involved in catalytic efficiency. (europa.eu)
THe OTCase lost most of its homotropic cooperativity and gained anabolic activity when residue glutamic acid (Glu) 106, near the CP binding site, was replaced by alanine (Ala) or glycine (Gly). (europa.eu)

Mechanisms of antimicrobial action and resistance in bacteria are complex and often vary depending on growth phase and/or concentration of the antimicrobial applied. (biorxiv.org)
Research applying genomic methods to develop treatments or vaccines, to understand antimicrobial mechanisms of resistance, and to examine evolutionary processes were also included. (cdc.gov)

Site-directed mutagenesis is a molecular biology method that is used to make specific and intentional mutating changes to the DNA sequence of a gene and any gene products. (wikipedia.org)
There are numerous methods for achieving site-directed mutagenesis, but with decreasing costs of oligonucleotide synthesis, artificial gene synthesis is now occasionally used as an alternative to site-directed mutagenesis. (wikipedia.org)
This synthetic primer contains the desired mutation and is complementary to the template DNA around the mutation site so it can hybridize with the DNA in the gene of interest. (wikipedia.org)
The gene thus copied contains the mutated site, and is then introduced into a host cell in a vector and cloned. (wikipedia.org)
The ProQ/FinO family of RNA binding proteins mediate sRNA-directed gene regulation throughout gram-negative bacteria. (nature.com)
instead, a convergence has taken place between conventional mutagenesis and NBTs, in particular due to the possibilities of TILLING methods that allow the fast detection of mutations in any gene of a genome. (frontiersin.org)
However, the differences between the strategies are 1) the precision of the exact site of mutation in a target gene, and 2) the number of off-target mutations affecting other genes than the target gene. (frontiersin.org)
This gene encodes an ubiquitously expressed, cytosolic form of TrxR, which functions as a homodimer containing FAD, and selenocysteine (Sec) at the active site. (cancerindex.org)

These methods of mutagenesis, however, are limited by the kind of mutation they can achieve, and they are not as specific as later site-directed mutagenesis methods. (wikipedia.org)
A large number of methods are available to effect site-directed mutagenesis, although most of them have rarely been used in laboratories since the early 2000s, as newer techniques allow for simpler and easier ways of introducing site-specific mutation into genes. (wikipedia.org)
Furthermore, an expression vector carrying the G1629R mutation was constructed by site-directed mutagenesis and transiently expressed in Cos-7 cells. (haematologica.org)
Mutation and in vitro DNA binding studies completed on a subset of the CRMs provide evidence for a direct regulatory relationship. (sdbonline.org)
Somatic mutation is considered the primary direct cause of tumorigenesis. (hep.com.cn)
The ZFN technique is a site-directed nuclease (SDN) tool that can be designed to produce a mutation at a predetermined position in the plant genome. (frontiersin.org)

The students will have opportunity to practice several of these methods during laboratory work, including site-directed mutagenesis, recombinant protein production and purification, real-time RT-PCR. (lu.se)

Although the signalling pathways induced by VEGF have been well-studied, the precise molecular mechanisms remain to be determined, particularly with respect to the relative contribution of the individual receptors. (figshare.com)
The molecular mechanisms remain elusive. (bvsalud.org)
Previous studies demonstrated that thiazides attenuate insulin secretion, but the molecular mechanisms remain elusive. (bvsalud.org)

Since 2013, the development of the CRISPR/Cas9 technology, based on a prokaryotic viral defense system, has also allowed for the editing of the genome, and mutagenesis may be performed in vivo with relative ease. (wikipedia.org)

We present data on the occurrence and characterization of the resistance mechanisms of Nal S -Cip DS isolates in 685 Salmonella Typhi isolates of the French National Reference Center for Salmonella (FNRC-Salm). (cdc.gov)

Site-directed mutagenesis is one of the most important laboratory techniques for creating DNA libraries by introducing mutations into DNA sequences. (wikipedia.org)
Knowledge on the details of the channel-binding sites of agitoxin 2 formed a basis for site-directed mutagenesis studies of the toxin and the K+ channel sequences. (rcsb.org)
Sequences of previously described binding sites were collected for these five factors present in the cis-regulatory regions of known target genes. (sdbonline.org)
The possible hypoxia response element (HRE) sequences in 5' non-coding Region before ATG start codon of Sema4D were screened, followed by point mutagenesis and luciferase assay analysis. (jcancer.org)

These studies provide the first crystallographic evidence of the mechanism by which nonsteroidal ligands interact with the wild type AR. (rcsb.org)
A molecular mechanism of ca- chemical reaction producing cyanate and elemental sulfur as final talysis is suggested based on high-resolution three-dimensional products (21, 22): structures, electron paramagnetic resonance (EPR) spectroscopy, quantum mechanics/molecular mechanics (QM/MM) simulations, - - 0 + - N ≡ C-S + H2O → N ≡ C-O + S + 2H + 2e [1] kinetic studies, and the results of site-directed mutagenesis. (docslib.org)
These structures, together with functional studies, reveal major conformational states of the NCC and an intriguing regulatory mechanism. (bvsalud.org)
Nevi may also develop on sun-protected sites, and studies regarding the efficacy of sunscreen in preventing nevus development have yielded conflicting results. (medscape.com)
Despite the widespread use of urea in studies of protein a combination of these mechanisms. (lu.se)
In this chapter, electrophilic agents nogenicity from studies of exposed (benzene, 1,3-butadiene, and eth- include direct-acting electrophilic humans. (who.int)
However, nevi are known to develop on sun-protected sites, and studies regarding the efficacy of sunscreen in preventing nevus development have yielded conflicting results. (medscape.com)
However, with an adeq battery of necf expsre blmarkers, prospective studies of en- vironmental effects on pregnancy outcomes might be possible. (cdc.gov)

Molecular biology of parasite and vector (with special emphasis to Plasmodium-Anopheles, Dengue-Aedes);Cellular and humoral mechanisms involved inimmunityto parasites;Immunomodulation in vectors;Vector parasite interactions and control strategies - Disease symptoms, Molecular approaches to diagnosis, Epidemiology, drug targets, vaccine strategies and proteomic approaches, prophylaxis, treatments to parasitic diseases and vector control measures, mechanisms of drug resistance in parasites or vectors. (bits-pilani.ac.in)

Currently, there is an incomplete understanding of the genetic basis for mechanisms underpinning survival in stressful conditions, such as the presence of anti-microbials. (biorxiv.org)

Several natural and synthetic small molecules specifically inhibit the Sec61 channel, generating cellular effects that are potentially useful for therapeutic purposes, but their inhibitory mechanisms remain unclear. (biorxiv.org)

Further work is now needed to investigate the exact molecular mechanisms behind the activation of p63. (elifesciences.org)
To investigate whether binding site clustering could help to explain the specificity of these factors for eve , a simple notion of binding site clustering was incorporated into CIS-ANALYST, allowing searches for segments of a specified length containing a minimum number of predicted binding sites. (sdbonline.org)
To investigate why subtle structural changes to both the ligand and the receptor (i.e. mutations) result in drastic changes in activity, we studied structure-activity relationships for nonsteroidal AR ligands through crystallography and site-directed mutagenesis, comparing bound conformations of R-bicalutamide, hydroxyflutamide, and two previously reported nonsteroidal androgens, S-1 and R-3. (rcsb.org)
To investigate possible mechanisms of compromised systemic immunity, we evaluated the ability of NP to directly affect stimulatory/polarizing activities of conventional DC (cDC) towards T cells in vitro. (cdc.gov)

He developed a structural model for the relevant calcium pump based upon its amino acid sequence and the properties of over 250 altered pumps created using the technique of site-directed mutagenesis. (royalsociety.org)

Badarau A, Dennison C. Copper trafficking mechanism of CXXC-containing domains: Insight from the pH-dependence of their Cu(I) afffinities . (ncl.ac.uk)

If this is the case, interactions, but it is not clear whether denaturants like urea then solvent denaturation can be driven simply by the ex- act directly by binding to the protein surface or indirectly by posure of more binding sites in the denatured protein perturbing solvent-mediated hydrophobic interactions or by (Schellman 1987). (lu.se)

Additionally, UV radiation contributes to direct DNA damage, the formation of reactive oxygen species, and the production of epidermal cytokines, growth factors, and neuropeptides. (medscape.com)
SCLC is a neuroendocrine carcinoma that exhibits aggressive behavior, rapid growth, early spread to distant sites, exquisite sensitivity to chemotherapy and radiation, and frequent association with distinct paraneoplastic syndromes , including hypercalcemia, Eaton-lambert syndrome, syndrome of inappropriate diuretic hormone, and many others. (medscape.com)

We welcome research that shows new or significantly improved protocols or methodologies in total synthesis, synthetic methodology or physical and theoretical organic chemistry as well as research that shows a significant advance in the organic chemistry or molecular design aspects of chemical biology, catalysis, supramolecular and macromolecular chemistry, theoretical chemistry, mechanism-oriented physical organic chemistry, medicinal chemistry or natural products. (rsc.org)

Find support for a specific problem in the support section of our website. (mdpi.com)
3) when the S1 bundle breaks apart leading to the fully open conformation, S1 helices may dock to a specific site on the inner surface of the pore. (usda.gov)
New Breeding Techniques (NBTs) include several new technologies for introduction of new variation into crop plants for plant breeding, in particular the methods that aim to make targeted mutagenesis at specific sites in the plant genome (NBT mutagenesis). (frontiersin.org)
We investigated the mechanism of neutralization of VEEV infection by the broadly cross-reactive E1-specific MAb 1A4B-6. (cdc.gov)

Battistuzzi G, Borsari M, Dennison C, Li C, Ranieri A, Sola M, Yanagisawa S. Active site loop dictates the thermodynamics of reduction and ligand protonation in cupredoxins . (ncl.ac.uk)

Direct interaction between PIP2 and a subset of class II PDZ domains (syntenin, CASK, Tiam-1) has been demonstrated. (embl.de)

These data revealed previously unreported interactions between VEGFRs and WW domain-containing proteins - interactions that could have important implications for the mechanisms regulating VEGF expression. (figshare.com)
TcDH contains a unique metal site comprising Crated into proteins for the purposes of copper trafficking three copper ions in a configuration that enables two-electron and storage (3, 4) and electron transport and catalysis (5-7). (docslib.org)
Its substrate specificity suggests that Tsp may contain a substrate recognition domain, which selectively binds to the nonpolar C-termini of substrate proteins, separate from its catalytic site. (embl.de)

By using published DNA binding specificity data for five transcription factors active in the early Drosophila embryo, genomic regions containing unusually high concentrations of predicted binding sites were identified for these factors. (sdbonline.org)
Using CIS-ANALYST, the distribution of Bcd, Cad, Hb, Kr, and Kni binding sites were examined in a 1-Mb genomic region surrounding the well-characterized eve locus at a site_p value of 0.0003. (sdbonline.org)

Site-directed mutagenesis was achieved in 1974 in the laboratory of Charles Weissmann using a nucleotide analogue N4-hydroxycytidine, which induces transition of GC to AT. (wikipedia.org)

In addition, these two binding sites play different roles in the regulation of Sema4D expression in HUVEC and Caco-2 cells. (jcancer.org)
Regulation of a carbon concentrating mechanism through CCM1 in Chlamydomonas reinhardtii. (bio.net)

Observed interactions between mutated toxin and channel are being used to elucidate the channel structure and mechanisms of channel-toxin interactions. (rcsb.org)
Common among these 10 interactions of organic compounds, their power to detect excess cancer agents is the electrophilic nature of or their metabolites, with DNA and R 1 risk at particular sites. (who.int)

Many approaches have since been developed to improve the efficiency of mutagenesis. (wikipedia.org)

A common feature of these cis-regulatory modules is the presence of multiple binding sites for multiple transcription factors. (sdbonline.org)
Our study describes the sequence characteristics of 5' non-coding region of Sema4D, enhances our understanding of the regulatory mechanism of Sema4D and benefits the development of a possible anti-angiogenesis therapeutic strategy for malignancies. (jcancer.org)

The presence of the disulfide bridge isolates the active site from solvent and hinders the binding of PLP and glycine in the active site. (nih.gov)

These data indicate that opioid and non-opioid mechanisms come into play under different circumstances. (jneurosci.org)
Macedo S, Pechlaner M, Schmid W, Weik M, Sato K, Dennison C, Djinovic-Carugo K. Can soaked-in scavengers protect metalloprotein active sites from reduction during data collection? . (ncl.ac.uk)
Conclusions AlbaTraDIS provides a simple and rapid method to analyse multiple transposon mutagenesis data sets allowing this technology to be used at large scale. (biorxiv.org)
Oral and pharyngeal cancers were es- there were discrepancies between ondary and unspecified sites (C76- timated to affect more than 482 000 data sources, the data in the hard files C80) were erroneously registered as people globally in 2008, more than half were used. (who.int)

Where methods are directed towards a narrow range of structures, the importance of these targets must be clearly justified. (rsc.org)
Following its decision, the EU Court considers that the NBTs operate "at a rate out of all proportion to those resulting from the application of conventional methods of mutagenesis. (frontiersin.org)

The mechanism of activation of the transient receptor potential vanilloid 4 (TRPV4) channel by 4α-phorbol esters was investigated by combining information from chemical modification of 4α-phorbol-didecanoate (4α-PDD, 2a ), site-directed mutagenesis, Ca 2+ imaging, and electrophysiology. (acs.org)
The decision was based in a context of conventional plant breeding using mutagenesis of crop plants by physical or chemical treatments. (frontiersin.org)
Due to its long safety record, organisms obtained by physical and chemical mutagenesis are exempt from the provisions of the GMO legislation in the EU. (frontiersin.org)

at more restrictive values, many of these sites would be lost (Berman, 2002). (sdbonline.org)
The same clustering criteria that were successful for identifying CRMs in eve (700-bp regions with at least 13 predicted binding sites) identified clusters overlapping 14 of these 19 known CRMs (Berman, 2002). (sdbonline.org)

The theoretical part covers the molecular principles and mechanisms underlying the experimental approaches. (lu.se)

The placebo effect is a psychobiological phenomenon that can be attributable to different mechanisms, including expectation of clinical improvement and pavlovian conditioning. (jneurosci.org)
See Small Cell Lung Cancer: Beating the Spread , a Critical Images slideshow, to help identify the key clinical and biologic characteristics of small cell lung cancer, the staging criteria, and the common sites of spread. (medscape.com)

Guzzi R, Sportelli L, Yanagisawa S, Li C, Kostrz D, Dennison C. The influence of active site loop mutations on the thermal stability of azurin from Pseudomonas aeruginosa . (ncl.ac.uk)
A fundamental understanding of protein stability and the mechanism of denaturant action must ultimately rest on detailed knowledge about the structure, solvation, and energetics of the denatured state. (lu.se)

U. Ryde, L. Hemmingsen (1997) "The active-site metal coordination geometry of cadmium-substituted alcohol dehydrogenase. (lu.se)

The crystal structure of the apoenzyme shows the C terminus locked in a closed conformation by a disulfide bond between Cys(972) in the C terminus and Cys(353) located in the active site. (nih.gov)
Reduction of the 353-972 disulfide releases the C terminus and allows access to the active site. (nih.gov)

The site variation rates of T471C/A600G/C862T were 72.7%, 18.2%, and 72.7% in cancer cells respectively. (jcancer.org)

Our website will not work properly. (rcsb.org)
Some features of this site may not work without it. (massey.ac.nz)

The observation of the disulfide bond that acts to stabilize the closed form suggests a molecular mechanism for the redox-dependent activation of glycine decarboxylase observed earlier. (nih.gov)
[ 15 ] Additionally, BRAF mutations have been detected in colorectal carcinomas and papillary thyroid carcinomas , which suggests that additional pathways for BRAF mutagenesis may exist. (medscape.com)

These findings explain the mechanism for thiazide-induced glucose intolerance and reveal a fundamental role of CA5b in TCA cycle anaplerosis and insulin secretion in ß cells. (bvsalud.org)

These results provide a novel mechanism whereby the SH2 domain of cten-mediated focal adhesion localization of DLC-1 plays an essential role in its tumor suppression activity. (rupress.org)
Our results provide critical insights for understanding the Na-Cl cotransport mechanism of the NCC, and they establish a framework for future drug design and for interpreting disease-related mutations. (bvsalud.org)

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