Interaction between the 67 kilodalton metastasis-associated laminin receptor and laminin.
Normal and neoplastic cells interact with laminin via a variety of cell surface proteins. The specific binding sites on laminin for each particular cell surface laminin-binding protein have not yet been identified. In this study, the interaction between laminin and the high affinity metastasis-associated 67 kD laminin receptor (67 LR) was investigated by electron microscopy using the rotary shadowing technique. Laminin receptor that was purified from human colon carcinoma metastases appeared as a globular structure with a diameter of 5.2 +/- 0.8 nm. The 67 LR specifically bound to laminin on its long arm close to the intersection of the long and the short arms. There was no specific interaction of bovine serum albumin with laminin. Biochemical confirmation of the rotary shadowing experiments included slot blot solid phase assays in which [I125]-labeled 67 LR bound in a dose dependent manner to laminin as well as to the chymotrypsin resistant (C1) fragment of laminin that contains a short piece of the long arm. [I125]-labeled 67 LR did not bind to the pepsin resistant (P1) fragment of laminin that did not contain that segment on the long arm. This study therefore identifies the binding site on laminin for the 67 kD metastasis-associated laminin receptor as a region on the long arm of laminin close to the intersection of the four arms.
Cioce
V
V
Tumor Invasion and Metastasis Section, National Cancer Institute, Bethesda, Maryland.
Margulies
I M
IM
Sobel
M E
ME
Castronovo
V
V
D000595:Amino Acid Sequence; D000818:Animals; D006801:Humans; D066298:In Vitro Techniques; D007797:Laminin / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D051379:Mice; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009362:Neoplasm Metastasis / Q000503:physiopathology*; D010446:Peptide Fragments / Q000378:metabolism; D011485:Protein Binding; D017474:Receptors, Laminin / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure
D007797:Laminin; D010446:Peptide Fragments; D017474:Receptors, Laminin