Related Articles19F NMR reveals the conformational properties of free thrombin and its zymogen precursor prethrombin-2.
J Biol Chem. 2020 May 01;:
Authors: Ruben EA, Gandhi PS, Chen Z, Koester SK, DeKoster GT, Frieden C, Di Cera E
Abstract
The conformational properties *of trypsin-like proteases and their zymogen forms remain controversial because of a lack of sufficient information on their free forms. Specifically, it is unclear whether the free protease is zymogen-like and shifts to its mature form upon a ligand induced fit or exists in multiple conformations in equilibrium from which the ligand selects the optimal fit via conformational selection. Here we report the results of 19F NMR measurements that reveal the conformational properties of a protease and its zymogen precursor in the free form. Using the trypsin-like, clotting protease thrombin as a relevant model system, we show that its conformation is quite different from that of its direct zymogen precursor prethrombin-2 and more similar to that of its fully active Na+-bound form. The results cast doubts on recent hypotheses that free thrombin is zymogen-like and transitions to protease-like forms upon ligand binding. Rather, they validate the scenario emerged from previous findings of X-ray crystallography and rapid kinetics supporting a pre-existing equilibrium between open (E) and closed (E*) forms of the active site. In this scenario, prethrombin-2 is more dynamic and exists predominantly in the E* form, whereas thrombin is more rigid and exists predominantly in the E form. Ligand binding to thrombin takes place exclusively in the E form without significant changes in the overall conformation. In summary, these results disclose the structural architecture of the fee forms of thrombin and prethrombin-2, consistent with an E*-E equilibrium and providing no evidence that free thrombin is zymogen-like.
PMID: 32358061 [PubMed - as supplied by publisher]
[NMR paper] In-cell NMR reveals potential precursor of toxic species from SOD1 fALS mutants.
In-cell NMR reveals potential precursor of toxic species from SOD1 fALS mutants.
In-cell NMR reveals potential precursor of toxic species from SOD1 fALS mutants.
Nat Commun. 2014;5:5502
Authors: Luchinat E, Barbieri L, Rubino JT, Kozyreva T, Cantini F, Banci L
Abstract
Mutations in the superoxide dismutase 1 (SOD1) gene are related to familial cases of amyotrophic lateral sclerosis (fALS). Here we exploit in-cell NMR to characterize the protein folding and maturation of a series of fALS-linked SOD1 mutants in human cells and to...
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[NMR paper] Cell-free Expressed Bacteriorhodopsin in Different Soluble Membrane Mimetics: Biophysical Properties and NMR Accessibility.
Cell-free Expressed Bacteriorhodopsin in Different Soluble Membrane Mimetics: Biophysical Properties and NMR Accessibility.
Cell-free Expressed Bacteriorhodopsin in Different Soluble Membrane Mimetics: Biophysical Properties and NMR Accessibility.
Structure. 2013 Feb 12;
Authors: Etzkorn M, Raschle T, Hagn F, Gelev V, Rice AJ, Walz T, Wagner G
Abstract
Selecting a suitable membrane-mimicking environment is of fundamental importance for the investigation of membrane proteins. Nonconventional surfactants, such as amphipathic polymers...
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[NMR paper] Conformational properties of peptide fragments homologous to the 106-114 and 106-126
Conformational properties of peptide fragments homologous to the 106-114 and 106-126 residues of the human prion protein: a CD and NMR spectroscopic study.
Related Articles Conformational properties of peptide fragments homologous to the 106-114 and 106-126 residues of the human prion protein: a CD and NMR spectroscopic study.
Org Biomol Chem. 2005 Feb 7;3(3):490-7
Authors: Di Natale G, Impellizzeri G, Pappalardo G
Two peptide fragments, corresponding to the amino acid residues 106-126 (PrP) and 106-114 (PrP) of the human prion protein have...
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[NMR paper] NMR structure of free RGS4 reveals an induced conformational change upon binding Galp
NMR structure of free RGS4 reveals an induced conformational change upon binding Galpha.
Related Articles NMR structure of free RGS4 reveals an induced conformational change upon binding Galpha.
Biochemistry. 2000 Jun 20;39(24):7063-73
Authors: Moy FJ, Chanda PK, Cockett MI, Edris W, Jones PG, Mason K, Semus S, Powers R
Heterotrimeric guanine nucleotide-binding proteins (G-proteins) are transducers in many cellular transmembrane signaling systems where regulators of G-protein signaling (RGS) act as attenuators of the G-protein signal cascade...
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NMR resonance assignments of thrombin reveal the conformational and dynamic effects o
NMR resonance assignments of thrombin reveal the conformational and dynamic effects of ligation
Lechtenberg, B. C., Johnson, D. J. D., Freund, S. M. V., Huntington, J. A....
The serine protease thrombin is generated from its zymogen prothrombin at the end of the coagulation cascade. Thrombin functions as...
Date: 2010-08-10
Source: PNAS
Number: 32
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Solid-State NMR Reveals Structural and Dynamical Properties of a Membrane Protein
http://pubs.acs.org/cgi-bin/abstract.cgi/jacsat/2007/129/i21/abs/ja069028m.html
Solid-State NMR Reveals Structural and Dynamical Properties of a Membrane-Anchored Electron-Carrier Protein, Cytochrome b<sub>5</sub>
<aui auinm="Durr, U. H. N."> <aui auinm="Yamamoto, K."> <aui auinm="Im, S.-C."> <aui auinm="Waskell, L."> <aui auinm="Ramamoorthy, A."> <aug><aul></aul></aug></aui></aui></aui></aui></aui> <au>Ulrich H. N. Dürr,</au> <au>Kazutoshi Yamamoto,</au><au>Sang-Choul Im,</au><au>Lucy Waskell,and </au><au>Ayyalusamy Ramamoorthy*</au>
*ramamoor@umich.edu
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19F NMR reveals the conformational properties of free thrombin and its zymogen precursor prethrombin-2.
Linear Mode
