Ping Hu1, Zhuan Bian, Mingwen Fan, Meijing Huang, Ping Zhang. 1. The Key Laboratory for Oral Biomedical Engineering of Ministry of Education, School and Hospital of Stomatology, Wuhan University, 237 Luoyu Road, Wuhan 430079, China. gloriaping@126.com
Abstract
INTRODUCTION: The general secretory (Sec) pathway is the main pathway for protein transport. Sortase A has been shown to anchor surface protein containing an LPXTG motif to the cell wall. In order to know the distribution pattern of the Sec translocons and sortase A in Streptococcus mutans, we explored the subcellular localisation of SecA and sortase A. METHODS: The anti-SecA antibody and anti-sortase A antibody were examined on Western blots to evaluate specificities. An indirect post-embedding immunogold method was used to determine the subcellular localisation of the Sec pathway and sortase A enzyme in S. mutans membrane. RESULTS: Immunoblotting experiments showed that the anti-SecA antibody and anti-sortase A antibody specifically recognised a single band of about 95 kDa and 27 kDa, respectively in the S. mutans GS-5 cell lysates. Immunogold electron micrographs showed that small and large gold particles clustered together at a distinct microdomain. CONCLUSIONS: These data showed for the first time that Sec translocase and sortase A colocalised to a single locus in the cytoplasmic membrane of S. mutans GS-5.
INTRODUCTION: The general secretory (Sec) pathway is the main pathway for protein transport. Sortase A has been shown to anchor surface protein containing an LPXTG motif to the cell wall. In order to know the distribution pattern of the Sec translocons and sortase A in Streptococcus mutans, we explored the subcellular localisation of SecA and sortase A. METHODS: The anti-SecA antibody and anti-sortase A antibody were examined on Western blots to evaluate specificities. An indirect post-embedding immunogold method was used to determine the subcellular localisation of the Sec pathway and sortase A enzyme in S. mutans membrane. RESULTS: Immunoblotting experiments showed that the anti-SecA antibody and anti-sortase A antibody specifically recognised a single band of about 95 kDa and 27 kDa, respectively in the S. mutans GS-5 cell lysates. Immunogold electron micrographs showed that small and large gold particles clustered together at a distinct microdomain. CONCLUSIONS: These data showed for the first time that Sec translocase and sortase A colocalised to a single locus in the cytoplasmic membrane of S. mutans GS-5.
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