New insight into transmembrane topology of Staphylococcus aureus histidine kinase AgrC

Biochim Biophys Acta. 2014 Mar;1838(3):988-93. doi: 10.1016/j.bbamem.2013.12.006. Epub 2013 Dec 17.

Abstract

Staphylococcus aureus accessory gene regulator (agr) locus controls the expression of virulence factors through a classical two-component signal transduction system that consists of a receptor histidine protein kinase AgrC and a cytoplasmic response regulator AgrA. An autoinducing peptide (AIP) encoded by agr locus activates AgrC, which transduces extracellular signals into the cytoplasm. Despite extensive investigations to identify AgrC-AIP interaction sites, precise signal recognition mechanisms remain unknown. This study aims to clarify the membrane topology of AgrC by applying the green fluorescent protein (GFP) fusion technique and the substituted cysteine accessibility method (SCAM). However, our findings were inconsistent with profile obtained previously by alkaline phosphatase. We report the topology of AgrC shows seven transmembrane segments, a periplasmic N-terminus, and a cytoplasmic C-terminus.

Keywords: AgrC; GFP; SCAM; Staphylococcus aureus; Topology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Amino Acid Substitution
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / metabolism*
  • Blotting, Western
  • Cell Membrane / metabolism*
  • Cysteine / metabolism*
  • Fluorescence
  • Green Fluorescent Proteins / metabolism*
  • Molecular Sequence Data
  • Phosphorylation
  • Protein Binding
  • Protein Kinases / chemistry*
  • Protein Kinases / metabolism*

Substances

  • Bacterial Proteins
  • Green Fluorescent Proteins
  • Protein Kinases
  • AgrC protein, Staphylococcus
  • Cysteine