Ligand and Redox Partner Binding Generates a New Conformational State in Cytochrome P450cam (CYP101A1)

2019 ◽  
Vol 141 (6) ◽  
pp. 2678-2683 ◽  
Author(s):  
Alec H. Follmer ◽  
Sarvind Tripathi ◽  
Thomas L. Poulos
Keyword(s):  
Conformational State ◽  
Cytochrome P450cam ◽  
Redox Partner

scholarly journals Delicate conformational balance of the redox enzyme cytochrome P450cam

2015 ◽  
Vol 112 (29) ◽  
pp. 9022-9027 ◽  
Author(s):  
Simon P. Skinner ◽  
Wei-Min Liu ◽  
Yoshitaka Hiruma ◽  
Monika Timmer ◽  
Anneloes Blok ◽  
...  
Keyword(s):  
Active Site ◽  
Paramagnetic Nmr ◽  
Energy Landscapes ◽  
Heme Group ◽  
Cytochrome P450cam ◽  
Redox Enzyme ◽  
Closed State ◽  
Redox Partner ◽  
Chemical Conditions ◽  
Physical And Chemical

The energy landscapes of proteins are highly complex and can be influenced by changes in physical and chemical conditions under which the protein is studied. The redox enzyme cytochrome P450cam undergoes a multistep catalytic cycle wherein two electrons are transferred to the heme group and the enzyme visits several conformational states. Using paramagnetic NMR spectroscopy with a lanthanoid tag, we show that the enzyme bound to its redox partner, putidaredoxin, is in a closed state at ambient temperature in solution. This result contrasts with recent crystal structures of the complex, which suggest that the enzyme opens up when bound to its partner. The closed state supports a model of catalysis in which the substrate is locked in the active site pocket and the enzyme acts as an insulator for the reactive intermediates of the reaction.

scholarly journals An Intermediate Conformational State of Cytochrome P450cam-CN in Complex with Putidaredoxin

Biochemistry ◽  
2019 ◽  
Vol 58 (18) ◽  
pp. 2353-2361 ◽  
Author(s):  
Shih-Wei Chuo ◽  
Lee-Ping Wang ◽  
R. David Britt ◽  
David B. Goodin
Keyword(s):  
Conformational State ◽  
Cytochrome P450cam

Some Approaches to Viscometric Study of Chitosan in Acetic Acid Solution

2016 ◽  
Vol 10 (2) ◽  
pp. 135-139 ◽  
Author(s):  
Valentina Chernova ◽  
◽  
Angela Shurshina ◽  
Elena Kulish ◽  
Gennady Zaikov ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Acid Solution ◽  
Intrinsic Viscosity ◽  
Acetic Acid Solution ◽  
Conformational State ◽  
Macromolecular Coil ◽  
Degree Of Swelling ◽  
Current Value ◽  
Viscometric Study

Some ways of estimating the values of the intrinsic viscosity of chitosan were analyzed. It was shown that the method of Irzhak and Baranov for estimating the current value of the intrinsic viscosity allows to adequately estimates the conformational state of the macromolecular coil and its degree of swelling.

scholarly journals Crystal structures of an E1–E2–ubiquitin thioester mimetic reveal molecular mechanisms of transthioesterification

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Lingmin Yuan ◽  
Zongyang Lv ◽  
Melanie J. Adams ◽  
Shaun K. Olsen
Keyword(s):  
Complex Formation ◽  
Crystal Structures ◽  
Molecular Mechanisms ◽  
Conformational State ◽  
Biochemical Data ◽  
Conformational States ◽  
Product Release ◽  
Closed Conformation ◽  
Open Conformation ◽  
Conjugating Enzymes

AbstractE1 enzymes function as gatekeepers of ubiquitin (Ub) signaling by catalyzing activation and transfer of Ub to tens of cognate E2 conjugating enzymes in a process called E1–E2 transthioesterification. The molecular mechanisms of transthioesterification and the overall architecture of the E1–E2–Ub complex during catalysis are unknown. Here, we determine the structure of a covalently trapped E1–E2–ubiquitin thioester mimetic. Two distinct architectures of the complex are observed, one in which the Ub thioester (Ub(t)) contacts E1 in an open conformation and another in which Ub(t) instead contacts E2 in a drastically different, closed conformation. Altogether our structural and biochemical data suggest that these two conformational states represent snapshots of the E1–E2–Ub complex pre- and post-thioester transfer, and are consistent with a model in which catalysis is enhanced by a Ub(t)-mediated affinity switch that drives the reaction forward by promoting productive complex formation or product release depending on the conformational state.

scholarly journals Human Lanosterol 14-Alpha Demethylase (CYP51A1) Is a Putative Target for Natural Flavonoid Luteolin 7,3′-Disulfate

Molecules ◽  
2021 ◽  
Vol 26 (8) ◽  
pp. 2237
Author(s):  
Leonid Kaluzhskiy ◽  
Pavel Ershov ◽  
Evgeniy Yablokov ◽  
Tatsiana Shkel ◽  
Irina Grabovec ◽  
...  
Keyword(s):  
Cholesterol Metabolism ◽  
Cholesterol Biosynthesis ◽  
Clinical Importance ◽  
Optical Biosensor ◽  
Water Soluble ◽  
Soluble Form ◽  
Redox Partner ◽  
Access Channel ◽  
Kd Values ◽  
Key Steps

Widespread pathologies such as atherosclerosis, metabolic syndrome and cancer are associated with dysregulation of sterol biosynthesis and metabolism. Cholesterol modulates the signaling pathways of neoplastic transformation and tumor progression. Lanosterol 14-alpha demethylase (cytochrome P450(51), CYP51A1) catalyzes one of the key steps in cholesterol biosynthesis. The fairly low somatic mutation frequency of CYP51A1, its druggability, as well as the possibility of interfering with cholesterol metabolism in cancer cells collectively suggest the clinical importance of CYP51A1. Here, we show that the natural flavonoid, luteolin 7,3′-disulfate, inhibits CYP51A1 activity. We also screened baicalein and luteolin, known to have antitumor activities and low toxicity, for their ability to interact with CYP51A1. The Kd values were estimated using both a surface plasmon resonance optical biosensor and spectral titration assays. Unexpectedly, in the enzymatic activity assays, only the water-soluble form of luteolin—luteolin 7,3′-disulfate—showed the ability to potently inhibit CYP51A1. Based on molecular docking, luteolin 7,3′-disulfate binding suggests blocking of the substrate access channel. However, an alternative site on the proximal surface where the redox partner binds cannot be excluded. Overall, flavonoids have the potential to inhibit the activity of human CYP51A1 and should be further explored for their cholesterol-lowering and anti-cancer activity.

scholarly journals Large domain movements upon UvrD dimerization and helicase activation

2017 ◽  
Vol 114 (46) ◽  
pp. 12178-12183 ◽  
Author(s):  
Binh Nguyen ◽  
Yerdos Ordabayev ◽  
Joshua E. Sokoloski ◽  
Elizabeth Weiland ◽  
Timothy M. Lohman
Keyword(s):  
Escherichia Coli ◽  
Single Molecule ◽  
Self Assembly ◽  
Total Internal Reflection ◽  
Dna Helicase ◽  
Conformational State ◽  
Helicase Activity ◽  
Multiple Conformations ◽  
Dna Substrate

Escherichia coli UvrD DNA helicase functions in several DNA repair processes. As a monomer, UvrD can translocate rapidly and processively along ssDNA; however, the monomer is a poor helicase. To unwind duplex DNA in vitro, UvrD needs to be activated either by self-assembly to form a dimer or by interaction with an accessory protein. However, the mechanism of activation is not understood. UvrD can exist in multiple conformations associated with the rotational conformational state of its 2B subdomain, and its helicase activity has been correlated with a closed 2B conformation. Using single-molecule total internal reflection fluorescence microscopy, we examined the rotational conformational states of the 2B subdomain of fluorescently labeled UvrD and their rates of interconversion. We find that the 2B subdomain of the UvrD monomer can rotate between an open and closed conformation as well as two highly populated intermediate states. The binding of a DNA substrate shifts the 2B conformation of a labeled UvrD monomer to a more open state that shows no helicase activity. The binding of a second unlabeled UvrD shifts the 2B conformation of the labeled UvrD to a more closed state resulting in activation of helicase activity. Binding of a monomer of the structurally similar Escherichia coli Rep helicase does not elicit this effect. This indicates that the helicase activity of a UvrD dimer is promoted via direct interactions between UvrD subunits that affect the rotational conformational state of its 2B subdomain.

scholarly journals SERR Spectroelectrochemical Study of Cytochrome cd1 Nitrite Reductase Co-Immobilized with Physiological Redox Partner Cytochrome c552 on Biocompatible Metal Electrodes

PLoS ONE ◽  
2015 ◽  
Vol 10 (6) ◽  
pp. e0129940 ◽  
Author(s):  
Célia M. Silveira ◽  
Pedro O. Quintas ◽  
Isabel Moura ◽  
José J. G. Moura ◽  
Peter Hildebrandt ◽  
...  
Keyword(s):  
Nitrite Reductase ◽  
Metal Electrodes ◽  
Redox Partner ◽  
Cytochrome C552 ◽  
Cytochrome Cd1

The interaction of the 2′-OH group with the vicinal phosphate in ribonucleoside 3′-ethylphosphate drives the sugar-phosphate backbone into unique (S,ω−) conformational state

Tetrahedron ◽  
1995 ◽  
Vol 51 (43) ◽  
pp. 11775-11792 ◽  
Author(s):  
J. Plavec ◽  
C. Thibaudeau ◽  
G. Viswanadham ◽  
C. Sund ◽  
A. Sandström ◽  
...  
Keyword(s):  
Conformational State ◽  
Sugar Phosphate ◽  
Phosphate Backbone
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