Proteins, structure, function, and bioinformatics, 2020-11, Vol.88 (11), p.1394-1400
2020
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- Autor(en) / Beteiligte
- Titel
- Engineering the specificity of Streptococcus pyogenes sortase A by loop grafting
- Ist Teil von
- Proteins, structure, function, and bioinformatics, 2020-11, Vol.88 (11), p.1394-1400
- Ort / Verlag
- Hoboken, USA: John Wiley & Sons, Inc
- Erscheinungsjahr
- 2020
- Quelle
- Wiley Online Library All Journals
- Beschreibungen/Notizen
- Sortases are a group of enzymes displayed on the cell‐wall of Gram‐positive bacteria. They are responsible for the attachment of virulence factors onto the peptidoglycan in a transpeptidation reaction through recognition of a pentapeptide substrate. Most housekeeping sortases recognize one specific pentapeptide motif; however, Streptococcus pyogenes sortase A (SpSrtA WT) recognizes LPETG, LPETA and LPKLG motifs. Here, we examined SpSrtA's flexible substrate specificity by investigating the role of the β7/β8 loop in determining substrate specificity. We exchanged the β7/β8 loop in SpSrtA with corresponding β7/β8 loops from Staphylococcus aureus (SaSrtA WT) and Bacillus anthracis (BaSrtA WT). While the BaSrtA‐derived variant showed no enzymatic activity toward either LPETG or LPETA substrates, the activity of the SaSrtA‐derived mutant toward the LPETA substrate was completely abolished. Instead, the mutant had an improved activity toward LPETG, the preferred substrate of SaSrtA WT.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 0887-3585eISSN: 1097-0134DOI: 10.1002/prot.25958Titel-ID: cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_7586933
- Format
- –
- Schlagworte
- Amino Acid Motifs, Aminoacyltransferases - chemistry, Aminoacyltransferases - genetics, Aminoacyltransferases - metabolism, Bacillus anthracis - chemistry, Bacillus anthracis - enzymology, Bacterial Proteins - chemistry, Bacterial Proteins - genetics, Bacterial Proteins - metabolism, Binding Sites, Cloning, Molecular, computational analysis, Cysteine Endopeptidases - chemistry, Cysteine Endopeptidases - genetics, Cysteine Endopeptidases - metabolism, Enzymatic activity, Escherichia coli - genetics, Escherichia coli - metabolism, Gene Expression, Genetic Vectors - chemistry, Genetic Vectors - metabolism, loop swapping, Models, Molecular, Mutants, Oligopeptides - chemistry, Oligopeptides - metabolism, Peptidoglycans, Protein Binding, Protein Conformation, alpha-Helical, Protein Conformation, beta-Strand, protein engineering, Protein Engineering - methods, Protein Interaction Domains and Motifs, Recombinant Proteins - chemistry, Recombinant Proteins - genetics, Recombinant Proteins - metabolism, Sequence Alignment, Sequence Homology, Amino Acid, Sortase, Staphylococcus aureus - chemistry, Staphylococcus aureus - enzymology, Streptococcus infections, Streptococcus pyogenes, Streptococcus pyogenes - chemistry, Streptococcus pyogenes - enzymology, Streptococcus pyogenes sortase A, Substrate Specificity, Substrates, Virulence, Virulence factors