Vlliv cysteinů na aktivitu a strukturu 12kDa formy proteasy M-PMV

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60461373%3A22330%2F05%3A00015783" target="_blank" >RIV/60461373:22330/05:00015783 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Affect of cysteines on the activity and structure of 12 kda form of m-pmv protease

Popis výsledku v původním jazyce

Mason-Pfizer monkey virus encodes an aspartic protease (M-PMV PR), which is essential for the correct assembly and maturation of the viral particles. Primary sequence of the protease contains two cysteine residues (Cys7, Cys106) that has been replaced byalanines to stabilize the monomeric form of the 12 kDa protease for structural study by NMR spectroscopy [1]. However, it was demonstrated that cysteines were not indifferent amino acid residues and might have a modulatory affect on the activity of theprotease [2]. Here we present results of structural and biochemical study on the inactive mutants (D26N) of the 12 kDa form of M-PMV PR with either one (C7A, C106A) or both cysteine residues preserved. We demonstrate that replacement of both cysteines affects not only the stability of the protease but also the proteolytic activity of the enzyme. MALDI-TOF measurements of a digest of PRD26N and a light scattering analysis revealed the formation of an intramolecular disulphide bridge, whic

Název v anglickém jazyce

Affect of cysteines on the activity and structure of 12 kda form of m-pmv protease

Popis výsledku anglicky

Mason-Pfizer monkey virus encodes an aspartic protease (M-PMV PR), which is essential for the correct assembly and maturation of the viral particles. Primary sequence of the protease contains two cysteine residues (Cys7, Cys106) that has been replaced byalanines to stabilize the monomeric form of the 12 kDa protease for structural study by NMR spectroscopy [1]. However, it was demonstrated that cysteines were not indifferent amino acid residues and might have a modulatory affect on the activity of theprotease [2]. Here we present results of structural and biochemical study on the inactive mutants (D26N) of the 12 kDa form of M-PMV PR with either one (C7A, C106A) or both cysteine residues preserved. We demonstrate that replacement of both cysteines affects not only the stability of the protease but also the proteolytic activity of the enzyme. MALDI-TOF measurements of a digest of PRD26N and a light scattering analysis revealed the formation of an intramolecular disulphide bridge, whic

Druh

O - Ostatní výsledky

CEP obor

CE - Biochemie

OECD FORD obor

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Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2005

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů