Identification of N-glycosylation in prolyl endoprotease from Aspergillus niger and evaluation of the enzyme for its possible application in proteomics

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61989592%3A15310%2F09%3A00010321" target="_blank" >RIV/61989592:15310/09:00010321 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/60162694:G44__/09:00002182

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Identification of N-glycosylation in prolyl endoprotease from Aspergillus niger and evaluation of the enzyme for its possible application in proteomics

Popis výsledku v původním jazyce

AA prolyl endoprotease from Aspergillus niger was isolated from the commercial product Brewers Clarex to evaluate itspossible application in proteomics. A colorimetric assessment with phenol and sulfuric acid showed the presence of neutral sugars (9 % ofweight). The subsequent treatment with N-glycosidase F released a variety of high-mannose type N-glycans, which were successfully detected using MALDI-TOF MS.MALDI-TOF/TOF tandem MS analysis of glycopeptides from a tryptic digest of prolyl endoproteaseunraveled the identity of the N-glycosylation site in the primary structure. Spectrophotometric measurements demonstrated optimal activity at pH 4.0-4.5 and also high thermostability for the cleavage at the C-terminal part of proline residues. The enzymeacts upon proline and alanine residues, but there is an additional minor cleavage at some other residues. The digestion of a honeybee peptide comprising six proline residues (apidaecin 1A) led to the detection of specific peptides termin

Název v anglickém jazyce

Identification of N-glycosylation in prolyl endoprotease from Aspergillus niger and evaluation of the enzyme for its possible application in proteomics

Popis výsledku anglicky

AA prolyl endoprotease from Aspergillus niger was isolated from the commercial product Brewers Clarex to evaluate itspossible application in proteomics. A colorimetric assessment with phenol and sulfuric acid showed the presence of neutral sugars (9 % ofweight). The subsequent treatment with N-glycosidase F released a variety of high-mannose type N-glycans, which were successfully detected using MALDI-TOF MS.MALDI-TOF/TOF tandem MS analysis of glycopeptides from a tryptic digest of prolyl endoproteaseunraveled the identity of the N-glycosylation site in the primary structure. Spectrophotometric measurements demonstrated optimal activity at pH 4.0-4.5 and also high thermostability for the cleavage at the C-terminal part of proline residues. The enzymeacts upon proline and alanine residues, but there is an additional minor cleavage at some other residues. The digestion of a honeybee peptide comprising six proline residues (apidaecin 1A) led to the detection of specific peptides termin

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

CE - Biochemie

OECD FORD obor

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Projekt

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Návaznosti

Z - Vyzkumny zamer (s odkazem do CEZ)<br>S - Specificky vyzkum na vysokych skolach

Rok uplatnění

2009

Kód důvěrnosti údajů

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

Název periodika

Journal of Mass Spectrometry

ISSN

1076-5174

e-ISSN

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Svazek periodika

44

Číslo periodika v rámci svazku

11

Stát vydavatele periodika

DE - Spolková republika Německo

Počet stran výsledku

9

Strana od-do

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Kód UT WoS článku

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EID výsledku v databázi Scopus

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