Assignment of laminin heavy chains using the lectin Ricinus communis agglutinin-1.
Using high-resolution PAGE and Western-blotting techniques the lectin Ricinus communis agglutinin-1 (RCA-1) was tested for its ability to recognize laminin subunits from the mouse Engelbreth-Holm-Swarm (EHS) tumour and from bovine cardiac and skeletal muscle. Biotinylated RCA-1 recognized both the A and B chains of purified EHS-tumour laminin with a sensitivity comparable to anti-(EHS laminin) antibodies. In cardiac and skeletal muscle RCA-1 also recognized the B chains of laminin, together with a approximately 330 kDa RCA-1-binding glycoprotein that was undetectable in smooth muscle. This glycoprotein was not recognized by antibodies raised to laminin from the EHS tumour. Purification of the 330 kDa binding glycoprotein from skeletal muscle, using ion-exchange and lectin-affinity chromatography, revealed that in its native form, this glycoprotein is disulphide-bonded to the B chains of laminin. The demonstrated properties of the approximately 330 kDa RCA-1-binding glycoprotein are identical to those reported for the variant M chain of merosin which is known to replace the A chain in laminin from the extrasynaptic regions of skeletal muscle. These results establish that biotinylated RCA-1 can recognize A-, B- and M-chain subunits of laminin isoforms, and that, when used in conjunction with other techniques, they provide a useful method for the assignment of laminin heavy chains.
Wadsworth
J D
JD
Department of Paediatrics and Neonatal Medicine, Royal Postgraduate Medical School, Hammersmith Hospital, London, U.K.
Okuno
A
A
Strong
P N
PN
D000818:Animals; D015153:Blotting, Western; D002367:Ricinus communis; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D007797:Laminin / Q000032:analysis*; D037102:Lectins; D051379:Mice; D015335:Molecular Probes; D009132:Muscles / Q000737:chemistry; D037121:Plant Lectins; D010947:Plants, Toxic; D012513:Sarcoma, Experimental / Q000737:chemistry
D007797:Laminin; D037102:Lectins; D015335:Molecular Probes; D037121:Plant Lectins; C017707:Ricinus communis agglutinin-1