Medical Microbiology and Immunology

, Volume 196, Issue 1, pp 1–10 | Cite as

Streptococcus agalactiae CAMP factor binds to GPI-anchored proteins

  • Shenhui Lang
  • Jie Xue
  • Zhongwu Guo
  • Michael Palmer
Original Investigation

Abstract

CAMP factor (protein B) is a pore-forming protein secreted by Streptococcus agalactiae. It causes lysis of sheep red blood cells when these have been sensitized with staphylococcal sphingomyelinase. We here show that CAMP factor binds to GPI-anchored proteins, and that this interaction involves the carbohydrate core of the GPI-anchor. Enzymatic cleavage of GPI-anchors with phosphatidylinositol-specific phospholipase C strongly reduces the sensitivity of erythrocytes to CAMP factor. Incorporation of alkaline phosphatase, a model GPI-anchored protein, into liposome membranes renders the latter susceptible to permeabilization by CAMP factor. GPI-anchored proteins therefore function as cellular receptors for CAMP factor.

Abbreviations

BAC-SulfoNHS

Biotinamidohexanoic acid 3-sulfo-N-hydroxysulfosuccinimide ester

C16-GPI

6-O-{{6-O-[N-(l-prolyl-l-seryl)-2-aminoethyl]-phosphonato-α-d-mannopyranosyl}-(1,2)-O-(α-d-mannopyranosyl)-(1,6)-O-(α-d-mannopyranosyl)-(1,4)-O-(2-amino-2-deoxy-α-d-glucopyranosyl)}-2-O-hexadecanoyl-myo-inositol

CHAPS

3-[(3-Cholamido propyl)-dimethylammonio]-1-propanesu1fonate

DMSO

Dimethylsulfoxide

DPPC

1,2-Dipalmitoyl-sn-glycero-3-phosphocholine

GPI

Glycosylphosphatidylinositol

HBS

20 mM HEPES, 150 mM NaCl, 10 mM MgCl2, pH 7.5

PBS

20 mM potassium phosphate, 150 mM NaCl, pH 7.5

PI-PLC

Phosphatidylinositol-specific phospholipase C

PLAP

Placental alkaline phosphatase

PMSF

Phenylmethylsulfonylfluoride

SDS-PAGE

Sodium dodecylsulfate polyacrylamide gel electrophoresis

TBS

100 mM Tris, 0.9% NaCl, pH 7.5

TBS-Tween

TBS with 0.1% (v/v) Tween 20

TMB

3,3′,5,5′-Tetramethylbenzidine

Triton-HEPES

20 mM HEPES (pH 7.5) containing 0.06% Triton X-100

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Shenhui Lang
    • 1
  • Jie Xue
    • 2
  • Zhongwu Guo
    • 2
  • Michael Palmer
    • 1
  1. 1.Department of ChemistryUniversity of WaterlooWaterlooCanada
  2. 2.Department of ChemistryWayne State UniversityDetroitUSA

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