Engineering the specificity of Streptococcus pyogenes sortase A by loop grafting

Magdalena Wojcik, Kamil Szala, Ronald van Merkerk, Wim J. Quax*, Ykelien L. Boersma

*Bijbehorende auteur voor dit werk

OnderzoeksoutputAcademicpeer review

2 Citaten (Scopus)
23 Downloads (Pure)

Samenvatting

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.

Originele taal-2English
Pagina's (van-tot)1394-1400
Aantal pagina's7
TijdschriftProteins
Volume88
Nummer van het tijdschrift11
Vroegere onlinedatum5-jun-2020
DOI's
StatusPublished - nov-2020

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