Abstract
Fibronectin binding domain was expressed on the cell surface of Lactobacillus casei strain Shirota which hardly adheres to fibronectin. DNA for the fibronectin binding domain of the sfbl gene, which encodes a fibronectin binding protein of Streptococcus pyogenes ATCC 21059, was amplified with polymerase chain reaction, cloned into a surface display vector pSAK332, and introduced into L. casei. The fibronectin binding domain was expressed as a fusion protein consisting of staphylokinase of Staphylococcus aureus and the anchor sequence of cell wall-associated 763 proteinase of Lactococcus lactis NCDO 763. The fibronectin binding ability of the resulting L. casei was confirmed with Western blot analysis, immunoelectron microscopic analysis, and adherence to fibroblast cells. These results indicate that L. casei has acquired a new phenotype to bind fibronectin upon the expression of the fibronectin binding domain on the cell surface. This L. casei also shows binding affinity to fibrinogen, indicating that fibronectin binding domain is involved in the binding to fibrinogen as well.
MeSH terms
-
Adhesins, Bacterial*
-
Amino Acid Sequence
-
Animals
-
Bacterial Adhesion
-
Bacterial Proteins / chemistry
-
Bacterial Proteins / genetics
-
Bacterial Proteins / metabolism
-
Base Sequence
-
Binding Sites / genetics
-
Carrier Proteins / chemistry
-
Carrier Proteins / genetics
-
Carrier Proteins / metabolism
-
Cell Line
-
Cell Membrane / metabolism
-
DNA, Bacterial / genetics
-
Fibrinogen / metabolism
-
Fibronectins / metabolism*
-
Lacticaseibacillus casei / genetics
-
Lacticaseibacillus casei / metabolism*
-
Lacticaseibacillus casei / ultrastructure
-
Metalloendopeptidases / chemistry
-
Metalloendopeptidases / genetics
-
Metalloendopeptidases / metabolism
-
Mice
-
Microscopy, Immunoelectron
-
Molecular Sequence Data
-
Phenotype
-
Protein Structure, Tertiary
-
Recombinant Fusion Proteins / chemistry
-
Recombinant Fusion Proteins / genetics
-
Recombinant Fusion Proteins / metabolism
-
Staphylococcus aureus / genetics
-
Staphylococcus aureus / metabolism
-
Streptococcus pyogenes / genetics
-
Streptococcus pyogenes / metabolism*
Substances
-
Adhesins, Bacterial
-
Bacterial Proteins
-
Carrier Proteins
-
DNA, Bacterial
-
Fibronectins
-
Recombinant Fusion Proteins
-
fibronectin-binding proteins, bacterial
-
Fibrinogen
-
Metalloendopeptidases
-
auR protein, Staphylococcus aureus