Single mutation on the surface of Staphylococcus aureus Sortase A can disrupt its dimerization

Biochemistry
Jie ZhuZhiwen Zhang

Abstract

Staphylococcus aureus Sortase A (SrtA) is an important Gram-positive membrane enzyme which catalyzes the anchoring of many cell surface proteins conserved with the LPXTG sequence. Recently SrtA has been demonstrated to be a dimer with a Kd of 55 microM in vitro. Herein, we show that a single point mutation of amino acid residue on the surface of SrtA can completely disrupt the dimerization. Native polyacrylamide gel electrophoresis and analytical gel filtration chromatography were used to detect the dimer-monomer equilibrium of SrtA mutants. Circular dichroism spectrum experiments were performed to study the conformational change of each SrtA mutant. An enzyme activity assay confirmed that all the SrtA mutants were active in vitro. Our results not only are important for understanding the SrtA protein self-associating mechanism but also provided the necessary starting materials for the study of sortase A pathway in vivo, which may have significant implications for discovering microbial physiology and give a potential target for novel Gram-positive antibiotics.

References

Jan 6, 1981·Biochemistry·S W Provencher, J Glöckner
Aug 14, 1999·The Journal of Biological Chemistry·H Ton-That, O Schneewind
Mar 24, 2004·Trends in Microbiology·Gavin K Paterson, Timothy J Mitchell
May 1, 2004·The Journal of Biological Chemistry·Yinong ZongSthanam V L Narayana
Feb 8, 2005·Current Opinion in Microbiology·Ivo G Boneca
Jun 20, 2006·Biophysical Journal·Petras J Kundrotas, Emil Alexov
Nov 23, 2006·FEBS Letters·Marlyse BuissonWim P Burmeister

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Citations

Dec 30, 2008·Analytical Biochemistry·Gabrielle Nina ThibodeauxZhiwen Zhang
May 8, 2009·Chembiochem : a European Journal of Chemical Biology·Aiko UmedaZhiwen Jonathan Zhang
Oct 27, 2011·Molecular Microbiology·Thomas SpirigRobert T Clubb
Sep 7, 2016·Journal of Molecular Recognition : JMR·Naiyf S AlharbiAhmed S Alobaidi
Sep 7, 2016·Biochemical and Biophysical Research Communications·Jian ChenBeiwen Zheng
Nov 11, 2017·Chembiochem : a European Journal of Chemical Biology·Keyvan D NikghalbJohn M Antos

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