Pseudotrypsin: a little known trypsin proteoform

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61989592%3A15310%2F18%3A73590131" target="_blank" >RIV/61989592:15310/18:73590131 - isvavai.cz</a>

Výsledek na webu

<a href="https://www.mdpi.com/1420-3049/23/10/2637" target="_blank" >https://www.mdpi.com/1420-3049/23/10/2637</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.3390/molecules23102637" target="_blank" >10.3390/molecules23102637</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Pseudotrypsin: a little known trypsin proteoform

Popis výsledku v původním jazyce

Trypsin is the protease of choice for protein sample digestion in proteomics. The most typical active forms are the single-chain beta-trypsin and the two-chain alfa-trypsin, which is produced by a limited autolysis of beta-trypsin. An additional intra-chain split leads to pseudotrypsin (psi-trypsin) with three chains interconnected by disulfide bonds, which can be isolated from the autolyzate by ion-exchange chromatography. Based on experimental data with artificial substrates, peptides, and protein standards, psi-trypsin shows altered kinetic properties, thermodynamic stability and cleavage site preference (and partly also cleavage specificity) compared to the above-mentioned proteoforms. In our laboratory, we have analyzed the performance of bovine psi-trypsin in the digestion of protein samples with a different complexity. It cleaves predominantly at the characteristic trypsin cleavage sites. However, in a comparison with common tryptic digestion, non-specific cleavages occur more frequently (mostly after the aromatic residues of Tyr and Phe) and more missed cleavages are generated. Because of the preferential cleavages after the basic residues and more developed side specificity, which is not expected to occur for the major trypsin forms (but may appear anyway because of their autolysis), psi-trypsin produces valuable information, which is complementary in part to data based on a strictly specific trypsin digestion and thus can be unnoticed following common proteomics protocols.

Název v anglickém jazyce

Pseudotrypsin: a little known trypsin proteoform

Popis výsledku anglicky

Trypsin is the protease of choice for protein sample digestion in proteomics. The most typical active forms are the single-chain beta-trypsin and the two-chain alfa-trypsin, which is produced by a limited autolysis of beta-trypsin. An additional intra-chain split leads to pseudotrypsin (psi-trypsin) with three chains interconnected by disulfide bonds, which can be isolated from the autolyzate by ion-exchange chromatography. Based on experimental data with artificial substrates, peptides, and protein standards, psi-trypsin shows altered kinetic properties, thermodynamic stability and cleavage site preference (and partly also cleavage specificity) compared to the above-mentioned proteoforms. In our laboratory, we have analyzed the performance of bovine psi-trypsin in the digestion of protein samples with a different complexity. It cleaves predominantly at the characteristic trypsin cleavage sites. However, in a comparison with common tryptic digestion, non-specific cleavages occur more frequently (mostly after the aromatic residues of Tyr and Phe) and more missed cleavages are generated. Because of the preferential cleavages after the basic residues and more developed side specificity, which is not expected to occur for the major trypsin forms (but may appear anyway because of their autolysis), psi-trypsin produces valuable information, which is complementary in part to data based on a strictly specific trypsin digestion and thus can be unnoticed following common proteomics protocols.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

<a href="/cs/project/LO1204" target="_blank" >LO1204: Udržitelný rozvoj výzkumu v Centru regionu Haná</a><br>

Návaznosti

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

Rok uplatnění

2018

Kód důvěrnosti údajů

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

Název periodika

MOLECULES

ISSN

1420-3049

e-ISSN

—

Svazek periodika

23

Číslo periodika v rámci svazku

10

Stát vydavatele periodika

CH - Švýcarská konfederace

Počet stran výsledku

14

Strana od-do

"2637-1"-"2637-14"

Kód UT WoS článku

000451201400233

EID výsledku v databázi Scopus

2-s2.0-85054848644