Identification of sortase substrates by specificity profiling

Autoren

Lena Schmohl
Jan Bierlmeier
Nicolai von Kügelgen
Leonie Kurz
Pascal Reis
Fabian Barthels
Pia Mach
Mike Schutkowski
Christian Freund
Dirk Schwarzer

DOI

10.1016/j.bmc.2017.06.033

ISSN

0968-0896

Ausgabe der Veröffentlichung

18

Zeitschrift

Bioorganic & Medicinal Chemistry

Sprache

en

Paginierung

5002 - 5007

Datum der Veröffentlichung

2017

Status

Published

Herausgeber

Elsevier BV

Herausgeber URL

http://dx.doi.org/10.1016/j.bmc.2017.06.033

Datum der Datenerfassung

2020

Titel

Identification of sortase substrates by specificity profiling

Ausgabe der Zeitschrift

25

Data source: Crossref

Abstract

Sortases catalyze the attachment of surface proteins to the peptidoglycan layer of gram-positive bacteria and further represent powerful tools of protein chemistry. During catalysis sortases cleave a donor substrate containing the LPxTG (x=any amino acid) sorting motif under formation of an enzyme-bound thioester and ligate this intermediate to an acceptor protein containing an N-terminal glycine residue. In addition to the well-established sortase A of Staphylococcus aureus several homologs of this enzyme have been identified in the genomes of gram-positive bacteria. We have profiled the specificity of seven sortases of Staphylococci and Streptococci origin and observed that sortases of the latter class displayed a more relaxed specificity for donor and acceptor substrates than their Staphylococci counterparts. Streptococci sortases prefer an LPKLG donor substrate sequence compared to the canonical sorting motif LPKTG. These findings might facilitate the use of Streptococci sortases as tools of protein chemistry.

Addresses

Interfaculty Institute of Biochemistry, University of Tuebingen, Hoppe-Seyler-Str. 4, D-72076 Tuebingen, Germany.

Autoren

Lena Schmohl
Jan Bierlmeier
Nicolai von Kügelgen
Leonie Kurz
Pascal Reis
Fabian Barthels
Pia Mach
Mike Schutkowski
Christian Freund
Dirk Schwarzer

DOI

10.1016/j.bmc.2017.06.033

eISSN

1464-3391

Externe Identifier

PubMed Identifier: 28684010

Funding acknowledgements

Deutsche Forschungsgemeinschaft: FR1325/15-1
Deutsche Forschungsgemeinschaft: SCHW 1163/7-1

Open access

false

ISSN

0968-0896

Ausgabe der Veröffentlichung

18

Zeitschrift

Bioorganic & medicinal chemistry

Schlüsselwörter

Staphylococcus
Cysteine Endopeptidases
Aminoacyltransferases
Peptides
Bacterial Proteins
Recombinant Proteins
Chromatography, High Pressure Liquid
Spectrometry, Fluorescence
Amino Acid Sequence
Substrate Specificity
Mass Spectrometry

Sprache

eng

Medium

Print-Electronic

Online publication date

2017

Paginierung

5002 - 5007

Datum der Veröffentlichung

2017

Status

Published

Datum der Datenerfassung

2017

Titel

Identification of sortase substrates by specificity profiling.

Sub types

Research Support, Non-U.S. Gov't
Journal Article

Ausgabe der Zeitschrift

25

Data source: Europe PubMed Central

Abstract

Sortases catalyze the attachment of surface proteins to the peptidoglycan layer of gram-positive bacteria and further represent powerful tools of protein chemistry. During catalysis sortases cleave a donor substrate containing the LPxTG (x=any amino acid) sorting motif under formation of an enzyme-bound thioester and ligate this intermediate to an acceptor protein containing an N-terminal glycine residue. In addition to the well-established sortase A of Staphylococcus aureus several homologs of this enzyme have been identified in the genomes of gram-positive bacteria. We have profiled the specificity of seven sortases of Staphylococci and Streptococci origin and observed that sortases of the latter class displayed a more relaxed specificity for donor and acceptor substrates than their Staphylococci counterparts. Streptococci sortases prefer an LPKLG donor substrate sequence compared to the canonical sorting motif LPKTG. These findings might facilitate the use of Streptococci sortases as tools of protein chemistry.

Date of acceptance

2017

Autoren

Lena Schmohl
Jan Bierlmeier
Nicolai von Kügelgen
Leonie Kurz
Pascal Reis
Fabian Barthels
Pia Mach
Mike Schutkowski
Christian Freund
Dirk Schwarzer

Autoren-URL

https://www.ncbi.nlm.nih.gov/pubmed/28684010

DOI

10.1016/j.bmc.2017.06.033

eISSN

1464-3391

Ausgabe der Veröffentlichung

18

Zeitschrift

Bioorg Med Chem

Schlüsselwörter

Protein labeling
Sortase
Sortase-mediated ligation
Staphylococcus aureus
Streptococcus pyogenes
Amino Acid Sequence
Aminoacyltransferases
Bacterial Proteins
Chromatography, High Pressure Liquid
Cysteine Endopeptidases
Mass Spectrometry
Peptides
Recombinant Proteins
Spectrometry, Fluorescence
Staphylococcus
Substrate Specificity

Sprache

eng

Country

England

Paginierung

5002 - 5007

PII

S0968-0896(17)30738-1

Datum der Veröffentlichung

2017

Status

Published

Datum, an dem der Datensatz öffentlich gemacht wurde

2017

Titel

Identification of sortase substrates by specificity profiling.

Sub types

Journal Article
Research Support, Non-U.S. Gov't

Ausgabe der Zeitschrift

25

Data source: PubMed

Identification of sortase substrates by specificity profiling

Tools