Lactobacillus casei acquires the binding activity to fibronectin by the expression of the fibronectin binding domain of Streptococcus pyogenes on the cell surface.
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.
Kushiro
A
A
Yakult Central Institute for Microbiological Research, Kunitachi, Tokyo, Japan. akira-kushiro@yakult.co.jp
Takahashi
T
T
Asahara
T
T
Tsuji
H
H
Nomoto
K
K
Morotomi
M
M
D018829:Adhesins, Bacterial; D000595:Amino Acid Sequence; D000818:Animals; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D004269:DNA, Bacterial / Q000235:genetics; D005340:Fibrinogen / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism*; D007780:Lacticaseibacillus casei / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D008666:Metalloendopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051379:Mice; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D010641:Phenotype; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013211:Staphylococcus aureus / Q000235:genetics / Q000378:metabolism; D013297:Streptococcus pyogenes / Q000235:genetics / Q000378:metabolism*
D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; D002352:Carrier Proteins; D004269:DNA, Bacterial; D005353:Fibronectins; D011993:Recombinant Fusion Proteins; C078635:fibronectin-binding proteins, bacterial; D005340:Fibrinogen; D008666:Metalloendopeptidases; C022653:auR protein, Staphylococcus aureus