Engineering the specificity of Streptococcus pyogenes sortase A by loop grafting.
Proteins
; 88(11): 1394-1400, 2020 11.
Article
em En
| MEDLINE
| ID: mdl-32501594
ABSTRACT
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 ß7/ß8 loop in determining substrate specificity. We exchanged the ß7/ß8 loop in SpSrtA with corresponding ß7/ß8 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.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Contexto em Saúde:
3_ND
Base de dados:
MEDLINE
Assunto principal:
Oligopeptídeos
/
Staphylococcus aureus
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Streptococcus pyogenes
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Bacillus anthracis
/
Proteínas de Bactérias
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Cisteína Endopeptidases
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Engenharia de Proteínas
/
Aminoaciltransferases
Tipo de estudo:
Prognostic_studies
Idioma:
En
Revista:
Proteins
Ano de publicação:
2020
Tipo de documento:
Article