Related Experiment Videos
Isolation and characterisation of a vitronectin-binding surface protein from Staphylococcus aureus
O D Liang1, J I Flock, T Wadström
1Department of Medical Microbiology, University of Lund, Sweden.
Biochimica Et Biophysica Acta
|July 3, 1995
Summary
Researchers isolated a 60 kDa protein from Staphylococcus aureus V8 that binds vitronectin. This protein is likely identical to a previously identified heparan sulfate-binding protein, suggesting a dual-binding capability in this bacterial strain.
Area of Science:
- Microbiology
- Biochemistry
- Protein Science
Background:
- Staphylococcus aureus strain V8 exhibits vitronectin binding.
- A 60 kDa protein was previously suggested as a high-affinity vitronectin-binding protein.
Purpose of the Study:
- To isolate and characterize the 60 kDa vitronectin-binding protein from Staphylococcus aureus V8.
- To investigate the binding properties and potential identity of this protein.
Main Methods:
- Bacterial cell surface proteins were extracted using 1 M LiCl.
- Protein separation was achieved using FPLC Mono-Q chromatography.
- Vitronectin-binding activity was assessed using microtiter plate assays, SDS-PAGE, Western blot, and ligand blotting.
Main Results:
- A single 60 kDa protein band with vitronectin-binding activity was isolated.
- The isolated protein bound soluble vitronectin in Western blot experiments.
- Amino-terminal sequencing revealed similarity to a 60 kDa heparan sulfate-binding protein from the same strain, suggesting molecular identity.
Conclusions:
- The 60 kDa vitronectin-binding protein from Staphylococcus aureus V8 has been successfully isolated and characterized.
- Evidence strongly suggests this protein is identical to the previously identified 60 kDa heparan sulfate-binding protein.
- This indicates a single protein molecule on the bacterial surface capable of binding both vitronectin and heparan sulfate.