Crystallographic and spectroscopic analyses of three hinge-bending mutants of the photoactive yellow protein are described. Previous studies have identified Gly(47) and Gly(51) as possible hinge points in the structure of the protein, allowing backbone segments around the chromophore to undergo large concerted motions. We have designed, crystallized, and solved the structures of three mutants: G47S, G51S, and G47S/G51S. The protein dynamics of these mutants are significantly affected. Transitions in the photocycle, measured with laser induced transient absorption spectroscopy, show rates up to 6-fold different from the wild type protein and show an additive effect in the double mutant. Compared with the native structure, no significant conformational differences were observed in the structures of the mutant proteins. We conclude that the structural and dynamic integrity of the region around these mutations is of crucial importance to the photocycle and suggest that the hinge-bending properties ...
Femtosecond time resolved pump-probe protein X-ray crystallography requires highly accurate measurements of the photoinduced structure factor amplitude differences. In the case of femtosecond photolysis of single P63 crystals of the Photoactive Yellow Protein, it is shown that photochemical dynamics place a Emerging Photon Technologies for Chemical Dynamics
We use polarization-sensitive ultrafast infrared spectroscopy to derive excited state lifetimes, quantum yields and structural information on the chromophore of photoactive yellow protein and model co
Addison, Kiri, Heisler, Ismael A., Conyard, Jamie, Dixon, Tara, Bulman Page, Philip C. and Meech, Stephen R. (2013) Ultrafast excited state dynamics of the green fluorescent protein chromophore and its kindling fluorescent protein analogue. Faraday Discussions, 163. pp. 277-296. ISSN 1359-6640 Full text not available from this repository. (Request a copy ...
The adsorption of photoactive yellow protein (PYP) on a Au(111) surface and its fluorescence activity have been studied by electrochemical scanning tunneling microscopy (EC-STM) and. uorescence photometry. A stable, densely packed protein layer was observed after protein immobilization onto a Au(111) surface modified with a mixture of 3-mercaptopropanoic acid (3-MPA) and 11-mercaptoundecanoic acid (11-MUA) and subsequent formation of the amide bond with the use of N-hydroxysuccinimide and carbodiimide. Fluorescence photometry data indicate that covalent binding of PYP to the functionalized Au(111) surface does not interfere with the. uorescence properties of the native protein.. ...
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2D02: The Crystal Structure of the R52Q Mutant Demonstrates a Role for R52 in Chromophore pK(a) Regulation in Photoactive Yellow Protein
2D01: The Crystal Structure of the R52Q Mutant Demonstrates a Role for R52 in Chromophore pK(a) Regulation in Photoactive Yellow Protein
Photoactive yellow protein (PYP) is a member of the xanthopsin family of eubacterial blue-light photoreceptors. On absorption of light, PYP enters a photocycle that ultimately transduces the energy contained in a light signal into an altered biological response. Nanosecond time-resolved x-ray crystallography was used to determine the structure of the short-lived, red-shifted, intermediate state denoted [pR], which develops within 1 nanosecond after photoelectronic excitation of the chromophore of PYP by absorption of light. The resulting structural model demonstrates that the [pR] state possesses the cis conformation of the 4-hydroxyl cinnamic thioester chromophore, and that the process of trans to cis isomerization is accompanied by the specific formation of new hydrogen bonds that replace those broken upon excitation of the chromophore. Regions of flexibility that compose the chromophore-binding pocket serve to lower the activation energy barrier between the dark state, denoted pG, and [pR], ...
To understand how the protein and chromophore components of a light-sensing protein interact to create a light cycle, we performed time-resolved spectroscopy on site-directed mutants of photoactive yellow protein (PYP). Recently determined crystallographic structures of PYP in the ground and colorless I2 states allowed us to design mutants and to study their photosensing properties at the atomic level. We developed a system for rapid mutagenesis and heterologous bacterial expression for PYP apoprotein and generated holoprotein through formation of a covalent thioester linkage with the p-hydroxycinnamic acid chromophore as found in the native protein. Glu46, replaced by Gln, is buried in the active site and hydrogen bonds to the chromophores phenolate oxygen in the ground state. The Glu46Gln mutation shifted the ground state absorption maximum from 446 to 462 nm, indicating that the color of PYP can be fine-tuned by the alteration of hydrogen bonds. Arg52, which separates the active site from ...
Use the presented tutorial to explore the formation of another type of chromophore and see the different states of its evolution. In addition learn more about the production of orange and red fluorescent proteins, called the mFruits.
PAS domains are found in diverse proteins throughout all three kingdoms of life, where they apparently function in sensing and signal transduction. Although a wealth of useful sequence and functional information has become recently available, these data have not been integrated into a three-dimensional (3D) framework. The very early evolutionary development and diverse functions of PAS domains have made sequence analysis and modeling of this protein superfamily challenging. Limited sequence similarities between the approximately 50-residue PAS repeats and one region of the bacterial blue-light photosensor photoactive yellow protein (PYP), for which ground-state and light-activated crystallographic structures have been determined to high resolution, originally were identified in sequence searches using consensus sequence probes from PAS-containing proteins. Here, we found that by changing a few residues particular to PYP function, the modified PYP sequence probe also could select PAS protein ...
SWISS-MODEL Template Library (SMTL) entry for 1s1y.1. Photoactivated chromophore conformation in Photoactive Yellow Protein (E46Q mutant) from 10 microseconds to 3 milliseconds
Andrea Carter, Susan Spiller, Eve Isabelle Charbonneau, Paulina Delgado, Stephanie Kwong, Thomas Huser, Nathan Rockwell, and Clark Lagarias. We are currently studying a family of cyanobacteriochromes through the use of a bacterium called Thermosynechococcus elongatus, which is a member of a family of thermophilic bacteria that contain cyanobacteriochromes. Cyanobacteriochromes contain GAF domains related to phytochromes. We are currently working with five genes from the genome of T. elongatus with class II GAF domains. GAF domains are blue/green photoreversible. This means that the organism can absorb green and blue photons, and once it is excited above a certain threshold in either color, it will reverse back to absorbing the opposite color, blue or green respectively. The genes we worked with include tlr0924, tll0569, tll0899, tlr0911, and tlr1999. All of these genes have been successfully truncated to GAF domain only and some have been expressed. I specifically worked with tlr1999 gene. Our ...
There are generally two absorption bands, denoted A and B, associated with GFP. The A-state and B-state are identified with the neutral chromophore and anionic [i.e. deprotonated] chromophore, respectively. The corresponding spectra are similar to the black and grey curves shown above. The green spectrum above is for the Cl1Y substituted chromophore, which is close to pK_a matching, and is rather broad and intermediate between the A-state and B-state spectra. This is arguably because the proton is delocalised between the chromophore and neighbouring Asp amino acid ...
Visual pigments with a longer peak-absorption wavelength or an open chromophore-binding pocket are generally noisier, being more prone to spontaneous activity.
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Membrane environment and fluidity can modulate the dynamics and interactions of membrane proteins and can thereby strongly influence the function of cells and organisms in general. In this work, we demonstrate that trans-cis isomerization of lipophilic dyes is a useful parameter to monitor packaging and fluidity of biomembranes. Fluorescence fluctuations, generated by trans-cis isomerization of the thiocarbocyanine dye Merocyanine 540 (MC540), were first analyzed by fluorescence correlation spectroscopy (FCS) in different alcohol solutions. Similar isomerization kinetics of MC540 in lipid vesicles could then also be monitored, and the influence of lipid polarity, membrane curvature, and cholesterol content was investigated. While no influence of membrane curvature and lipid polarity could be observed, a clear decrease in the isomerization rates could be observed with increasing cholesterol contents in the vesicle membranes. Finally, procedures to spatially map photoinduced and thermal ...
Mono- and Stereopictres of 5.0 Angstrom coordination sphere of Bromine atom in PDB 1o3k: Elaborate Manifold Of Short Hydrogen Bond Arrays Mediating Binding of Active Site-Directed Serine Protease Inhibitors
Mono- and Stereopictres of 5.0 Angstrom coordination sphere of Bromine atom in PDB 1o2y: Elaborate Manifold Of Short Hydrogen Bond Arrays Mediating Binding of Active Site-Directed Serine Protease Inhibitors
The high-yield expression and purification of cytochrome nitrite reductase (ccNiR), and its own characterization by a variety of methods, notably Laue crystallography, is reported. reduced species, the electrons are distributed amongst the numerous hemes, rather than being localized on specific heme centers. The purified ccNiR yielded good quality crystals, with which the 2.59 ? resolution structure was solved at Slc4a1 room heat using the Laue diffraction method. The structure is similar to that of ccNiR, except in the region where in fact the enzyme interacts using its physiological electron donor (CymA regarding ccNiR, NrfB regarding the proteins). nitrite reductase, NrfA, Laue crystallography, UV/Vis spectropotentiometry, Proteins film voltammetry Launch Ammonia-nitrite interconversion, a significant area of the natural nitrogen cycle, is certainly completed by a number of bacterias within their respiratory procedure. In one path the ammonia oxidizing bacterias (AOB) such as for example make ...
hort hydrogen bonds are present in many chemical and biological sys-tems. It is well known that these short hydrogen bonds are found in the active site of enzymes and aid enzyme catalysis. This study aims to system-atically characterize all short hydrogen bonds from a nonredundant dataset of protein structures. The study has revealed that short hydrogen bonds are commonly found in proteins and are widely present in different regions of the protein chain, such as the backbone or side chain, and in different secondary structural regions such as helices, strands and turns. ... ...
Rhodopsin is the protein found in retinas that converts light into a molecular signal that is eventually transmitted to the brain as a nerve signal. It is located in the disks in the outer segments of rod cells. When a photon is absorbed by the proteins chromophore, retinal, it undergoes a cis-trans conformational change. This structural change is carried out in the context of the protein structure and generates a change in structure on the cytoplasmic side of the protein than causes it to interact with the G-protein, transducin. A series of intermolecular interactions involving the Transducin alpha subunit and a phosphodiesterase eventually causes a reduction in the levels of c-GMP and the associated closing of ion channels in the rod cells. This causes the cells to polarize and starts the nervous signal. We have recently solved the three-dimensional structure of rhodopsin (Science, 289, 739-745 (2000)). Much work remains to structurally characterize the various photostates of the molecule and ...
We are developing femtosecond time-resolved impulsive Raman spectroscopy (femtosecond is 10-15 second), which enables tracking structural change of the reacting molecule through direct observation of the nuclear motion. Using this method, we successfully elucidated the first event occurring in a photoreceptor protein Yellow Protein (PYP) when it stimulates bacteria to escape from the harmful light under light irradiation. We first irradiated PYP with blue light and then shook PYP for observing the nuclear motion moment by moment, like taking stroboscopic photograph. It was revealed that only 10 trillionths of a second (10-13 seconds) from the irradiation of blue light, specific hydrogen bonds near the light-absorbing site weakened, and then carbon - carbon double bond rotated, generating a peculiar distorted structure. This is a breakthrough achievement that tracks elaborate mechanisms where proteins start to function in real time.. ...
In the course of their use as animal research models [more on this soon!], some axolotls got a pretty cool addition to their genomes: the GFP trait. Originally found in a species of jellyfish, this trait causes nearly every cell in the axolotls body to produce a bright yellow protein which glows neon green under a UV light. Why is this cool? First, its been very helpful to researchers working on limb regeneration and organ transplants. Second, it looks very pretty! And third, the trait can be passed down from generation to generation. But my favorite thing about it is that, since the effect isnt limited to pigment cells, it isnt affected by leucism. Youll see what I mean when we get to the next part!. ...
In the course of their use as animal research models [more on this soon!], some axolotls got a pretty cool addition to their genomes: the GFP trait. Originally found in a species of jellyfish, this trait causes nearly every cell in the axolotls body to produce a bright yellow protein which glows neon green under a UV light. Why is this cool? First, its been very helpful to researchers working on limb regeneration and organ transplants. Second, it looks very pretty! And third, the trait can be passed down from generation to generation. But my favorite thing about it is that, since the effect isnt limited to pigment cells, it isnt affected by leucism. Youll see what I mean when we get to the next part!. ...
Some experimental protocols, such as tandem affinity purification (TAP), generate complex data sets, depicting interactions in which more than two proteins are involved at the same time (n-ary interactions).. However, interaction data are often stored in tabular formats that aim to be amenable to quick, comprehensive searches. It may be desirable to convert these complexes into sets of binary interactions to simplify and speed up searches. There are two algorithms that will perform such conversion: the matrix model and the spoke model 2, as depicted in Figure 10. In this hypothetical example, take the bottom right protein complex (marked reality). A tandem affinity experiment (far left) might tell you that each of the other five proteins interact with the red bait protein in the middle. In reality, the red protein has only one interactor, which is the yellow protein. As you can see, both algorithms are somewhat mis-leading, but as the spoke model generates up to 3 times fewer false positives ...
|p|Audio seminar presented by Guy Finley at Life of Learning Foundation.|/p| |p||strong|Key Lesson:|/strong| Using thought as a tool to resolve the troubles that thought stirs up in the mind is like trying to use your fingers to seize, sort, and settle d
One of the primary rRNA binding proteins, it binds directly to 16S rRNA where it nucleates assembly of the body of the 30S subunit.
We report on gas-phase experiments investigating the predissociation and possible IR-driven isomerization of the 3-aminophenol-ammonia complex (3-AP-NH$_{3}$). A molecular beam of 3-AP-NH$_{3}$ is vibrationally excited with pulsed IR light, initiating an intramolecular vibrational redistribution and subsequent dissociation. The 3-AP fragment is then probed state-selectively via multiphoton ionization (REMPI) and time-of-flight mass spectrometry. Of particular interest is an IR-driven feature which we associate tentatively with a trans-cis isomerization process. We see clear correlation between the excitation of specific vibrational modes (namely the NH$_{3}$ symmetric and OH stretches) and the presence of this feature, as evidenced by IR-action and IR-depletion spectra. The feature persists atop a broader signal which we assign to the predissociation of the complex and whose cutoff in REMPI-action experiments provides an upper bound on the dissociation energy for 3-AP-NH$_{3 ...
Kang I, Johnson S, Lindenberg A, Falcone R, Missalla T, Heimann P, Kim KH, Katsufuji T, Cheong SW. 2001. Time-resolved x-ray measurements of polaron dynamics of charge-ordered Nd1/2Sr1/2MnO3. Ultrafast Phenomena XII. :287-289. ...
LIINA KANGUR. Tartu ülikool. Taotletav teaduskraad: filosoofiadoktor (PhD), biokeemia. Doktoritöö „Kõrgrõhu-spektroskoopiline uurimus fotosünteetiliste bakterite valgust koguvate valkude kromofooride vesiniksidemetest („High-pressure spectroscopy study of chromophore-binding hydrogen bonds in light-harvesting complexes of photosynthetic bacteria). hdl.handle.net/10062/31584. Kaitsmine toimub esmaspäeval, 26. augustil kl 10 Tartus Riia 23b Eesti biokeskuse auditooriumis nr 105.. Juhendaja: prof Arvi Freiberg. Oponent: prof Roland Winter (Dortmundi ülikool, Saksamaa). MAIDO HIIEMAA. Tallinna tehnikaülikool. Taotletav teaduskraad: filosoofiadoktor (PhD). Doktoritöö „Liikumise plaanija külisroolimisega mehitamata maasõidukile („Motion Planner for Skid-Steer Unmanned Ground Vehicle). http://digi.lib.ttu.ee/i/?932. Kaitsmine toimub esmaspäeval, 26. augustil kl 11 Tallinnas TTÜ VI õppehoone ruumis 442.. Juhendaja: prof Mart Tamre. Oponendid: prof Jānis Vība (Riia ...
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yellow labs - MedHelps yellow labs Center for Information, Symptoms, Resources, Treatments and Tools for yellow labs. Find yellow labs information, treatments for yellow labs and yellow labs symptoms.
안녕하세요 신박입니다 이번에는 카트라이더 이볼브 카트코드를 알려드리려고합니다 차량은 본카에 나온것도있지만 없는것도 많이있고 약을빨고 만든 카트도 몇몇 보이는거 같습니다 파일 다운로드 카트라이더.
Role of Mouse Cryptochrome Blue-Light Photoreceptor in Circadian Photoresponses. Does the p53 up-regulated Gadd45 protein have a role in excision repair?
TY - JOUR. T1 - Viscosity independence of the ratio of fluorescence lifetimes and quantum yields of ω-substituted 4- dimethylamino-trans-styrenes. AU - Kawski, Alfons. AU - Gryczynski, Ignacy. AU - Bojarski, Piotr. AU - Nowaczyk, Kazimierz. PY - 1992/9/15. Y1 - 1992/9/15. N2 - The fluorescence lifetimes τ and quantum yields φF of 4-dimethylamino-ω-diphenylphosphinyl- trans-styrene (2a) and 4-dimethylamino-ω-methylsulphonyl-trans-styrene (3a) were measured in n-paraffins (n = 6 to n = 16) at 293 K. The ratio τ/φF was found to be viscosity independent and equal to the radiative lifetime τ0′ calculated from the integrated absorption spectrum. The strong fluorescence quenching and the shortening of the lifetime τ when reducing the viscosity are due to twisting around the double bond, resulting in trans-cis isomerization.. AB - The fluorescence lifetimes τ and quantum yields φF of 4-dimethylamino-ω-diphenylphosphinyl- trans-styrene (2a) and ...
A Postdoctoral Research Position is available to study opsins and opsin-related proteins in the eukaryotic multicellular fungus Neurospora crassa. Our group was the first to clone, mutate and characterize an opsin from lower eukaryotes (See Bieszke et al, 1999, Proc. Natl. Acad. Sci. USA 96:8034-8039; Bieszke et al., 1999, Biochemistry 38:14138-14145). Possible projects include identification and characterization of additional opsins and opsin-related proteins, elucidation of the pathways regulated by these proteins, and determination of their chromophore-binding properties and photocycle characteristics. Applicants should have a Ph.D. degree in Biochemistry, Biology or a related discipline, have recombinant DNA experience and possess a good command of spoken and written English. Experience in microbiological techniques and protein biochemistry is desirable. Send a curriculum vitae (including a list of publications) and the names of three references to Dr. Katherine A. Borkovich, Associate ...
The course will be a study of the relevant federal laws concerning both public accounting and publicly traded companies, and the intersection of those laws with white collar fraud investigations. The course will also emphasize approaches to white collar investigations, including through the application of forensic accounting ...
White collar sign Signo de cuello blanco Separación radiolúcida entre la masa de coils y el lumen de la arteria a nivel del cuello. Este hallazgo radiotransparente (blanco), posiblemente, representa la formación de un tejido neointimal a nivel del cuello. La presencia de una señal de cuello blanco y la ausencia de
This site and any information contained including outbound links herein are intended for informational purposes only and should not be construed as legal advice. Seek competent legal counsel for advice on any legal matter. ...
Die Universität zu Köln ist eine Exzellenzuniversität mit dem klassischen Fächerspektrum einer Volluniversität. Als eine der größen Hochschulen Europas arbeitet sie in Forschung und Lehre auch international auf höchstem Niveau.
Dorn T, Kornherr J, Parrotta EI, Zawada D, Ayetey H, Santamaria G, Iop L, Mastantuono E, Sinnecker D, Goedel A, Dirschinger RJ, My I, Laue S, Bozoglu T, Baarlink C, Ziegler T, Graf E, Hinkel R, Cuda G, Kääb S, Grace AA, Grosse R, Kupatt C, Meitinger T, Smith AG, Laugwitz KL, Moretti A ...
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A group of microbial retinal proteins most closely related to the proton pump xanthorhodopsin has a novel sequence motif and a novel function. Instead of, or in addition to, proton transport, they perform light-driven sodium ion transport, as reported for one representative of this group (KR2) from Krokinobacter. In this paper, we examine a similar protein, GLR from Gillisia limnaea, expressed in Escherichia coli, which shares some properties with KR2 but transports only Na +. The absorption spectrum of GLR is insensitive to Na + at concentrations of ≤3 M. However, very low concentrations of Na + cause profound differences in the decay and rise time of photocycle intermediates, consistent with a switch from a Na +-independent to a Na +-dependent photocycle (or photocycle branch) at ~60 μM Na +. The rates of photocycle steps in the latter, but not the former, are linearly dependent on Na + concentration. This suggests that a high-affinity Na + binding site is created transiently after ...
Dark State continues the events of Charles Strosss previous novel, Empire Games, which is a combination of a science fiction and espionage thriller. Dark State continues the story of multiple timelines. Miriam Burgeson, one of the main characters, is the head of an espionage agency in Timeline Two that spies on the other three timelines. Timeline Two closely resembles our own history with the exception of a stolen nuclear bomb being detonated in the White House, killing the President of the United States. In Dark State, Timeline Three-the country called New American Commonwealth, which overthrew the British monarchy in 2003-is facing a succession crisis. The leader is terminally ill, and the timelines government is looking for a successor. As a result, the people who move between timelines-called World Walkers-who include Rita Douglas and Miriam Burgeson, are summoned to ensure that the New American Commonwealth of Timeline Three doesnt fall into chaos. Miriam and Rita must successfully ...
Phototaxis is one of the most fundamental stimulus-response behaviors in biology wherein motile microorganisms sense light gradients to swim toward the light source. Apart from single-cell survival and growth, it plays a major role at the global scale of aquatic ecosystems and bioreactors. We study phototaxis of single-celled algae Chlamydomonas reinhardtii as a function of cell number density and light stimulus using high spatiotemporal video microscopy. Surprisingly, the phototactic efficiency has a minimum at a well-defined number density, for a given light gradient, above which the phototaxis behavior of a collection of cells can even exceed the performance obtainable from single isolated cells. We show that the origin of enhancement of performance above the critical concentration lies in the slowing down of the cells, which enables them to sense light more effectively. We also show that this steady-state phenomenology is well captured by modeling the phototactic response as a ...
A group of microbial retinal proteins most closely related to the proton pump xanthorhodopsin has a novel sequence motif and a novel function. Instead of, or in addition to, proton transport, they perform light-driven sodium ion transport, as reported for one representative of this group (KR2) from Krokinobacter. In this paper, we examine a similar protein, GLR from Gillisia limnaea, expressed in Escherichia coli, which shares some properties with KR2 but transports only Na(+). The absorption spectrum of GLR is insensitive to Na(+) at concentrations of ≤3 M. However, very low concentrations of Na(+) cause profound differences in the decay and rise time of photocycle intermediates, consistent with a switch from a Na(+)-independent to a Na(+)-dependent photocycle (or photocycle branch) at ∼60 μM Na(+). The rates of photocycle steps in the latter, but not the former, are linearly dependent on Na(+) concentration. This suggests that a high-affinity Na(+) binding site is created transiently ...
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‎USA networks newest hit original series White Collar returns for a white-hot season two! White Collar, one of this fall seasons most critically acclaimed dramas, focuses on the most unlikely of partnerships between a con artist (Matt Bomer) and an FBI agent (Tim DeKay). Diahann Carro…
‎USA networks newest hit original series White Collar returns for a white-hot season two! White Collar, one of this fall seasons most critically acclaimed dramas, focuses on the most unlikely of partnerships between a con artist (Matt Bomer) and an FBI agent (Tim DeKay). Diahann Carro…
Name:C.I.Solvent Yellow 114,C.I.47020 Molecular Structure: Quinoline C.I.Solvent Yellow 114,C.I.47020,CAS 75216-45-4,7576-65-0,289.28,C18H11NO3,Solvent Yellow GS,Transparent Yellow 3G,Transparent Yellow G,Transparent Yellow HLR,Transparent Yellow S-2G,Oil Yellow G,Oil Yellow 3GE C.I.Solvent Yellow 114,C.I.47020,CAS 75216-45-4,7576-65-0,289.28,C18H11NO3,Solvent Yellow GS,Transparent Yellow 3G,Transparent Yellow G,Transparent Yellow HLR,Transparent Yellow S-2G,Oil Yellow G,Oil Yellow 3GE Molecular Formula:C18H11NO3 Molecular Weight: 289.28 CAS Registry Number:75216-45-4/7576-65-0
Lunevich A.Ya; Khmel'nitskii A.I.; Cherenkevich S.N.; Komyak A.I.; Min'ko A.A., 1983: Spectral fluorescent investigation of chromophores forming in the course of lipid per oxidation
The wave function of the ground state of a particle in a one-dimensional box is a half-period sine wave, which goes to zero at the two edges of the well. … This function is known as the 1s atomic orbital. For hydrogen (H), an electron in the ground state has energy −13.6 eV, relative to the ionization threshold.. ...