Blood platelets contain and secrete laminin-8 (alpha4beta1gamma1) and adhere to laminin-8 via alpha6beta1 integrin.
Laminins, a family of heterotrimeric proteins with cell adhesive/signaling properties, are characteristic components of basement membranes of vasculature and tissues. In the present study, permeabilized platelets were found to react with a monoclonal antibody to laminin gamma1 chain by immunofluorescence. In Western blot analysis of platelet lysates, several monoclonal antibodies to gamma1 and beta1 laminin chains recognized 220- to 230-kDa polypeptides, under reducing conditions, and a structure with much slower electrophoretic mobility under nonreducing conditions. Immunoaffinity purification on a laminin beta1 antibody-Sepharose column yielded polypeptides of 230, 220, 200, and 180 kDa from platelet lysates. In the purified material, mAbs to beta1 and gamma1 reacted with the two larger polypeptides, while affinity-purified rabbit antibodies to laminin alpha4 chain recognized the smallest polypeptide. Identity of the polypeptides was confirmed by microsequencing. One million platelets contained on average 1 ng of laminin (approximately 700 molecules per cell), of which 20-35% was secreted within minutes after stimulation with either thrombin or phorbol ester. Platelets adhered to plastic surfaces coated with the purified platelet laminin, and this process was largely inhibited by antibodies to beta1 and alpha6 integrin chains. We conclude that platelets contain and, following activation, secrete laminin-8 (alpha4beta1gamma1) and that the cells adhere to the protein by using alpha6beta1 integrin.
Geberhiwot
T
T
Microbiology and Tumorbiology Center, Karolinska Institutet, Stockholm, S 171 77, Sweden.
Ingerpuu
S
S
Pedraza
C
C
Neira
M
M
Lehto
U
U
Virtanen
I
I
Kortesmaa
J
J
Tryggvason
K
K
Engvall
E
E
Patarroyo
M
M
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001792:Blood Platelets / Q000737:chemistry* / Q000378:metabolism*; D015153:Blotting, Western; D005434:Flow Cytometry; D006801:Humans; D039121:Integrin alpha6beta1; D016023:Integrins / Q000276:immunology / Q000378:metabolism*; D007797:Laminin / Q000032:analysis / Q000378:metabolism*; D008969:Molecular Sequence Data; D010973:Platelet Adhesiveness / Q000502:physiology*
D000911:Antibodies, Monoclonal; D039121:Integrin alpha6beta1; D016023:Integrins; D007797:Laminin; C110308:laminin 8