Lectins as probes of chromatin structure. Binding of concanavalin A to purified rat liver chromatin.
Concanavalin A (Con A) binds specifically to rat liver chromatin. The extent of binding is directly proportional to both chromatin and concanavalin A concentration. It is reversible and inhibited by specific sugars for which concanavalin A has a binding site. Scatchard analysis reveals the presence of one type of Con A-binding site, with an apparent dissociation constant of 3 X 10(-7) M. A maximum of 10 pmol of Con A binds to 10 microgram of chromatin, indicating an average of one binding site/1400 base pairs of DNA. To identify the polypeptide chains which contain Con A-binding sites, chromosomal proteins were separated by electrophoresis on polyacrylamide gels containing sodium dodecyl sulfate. Con A receptors were localized by incubating the gel in 125I-Con A and subsequent autoradiography. Three major polypeptide bands which bind Con A were identified among the nonhistone chromosomal proteins. The apparent molecular weights of these glycoproteins are 135,000, 125,000, and 69,000. We suggest that lectins may serve as probes for the study of the organization of specific components in chromatin.
D000818:Animals; D002843:Chromatin / Q000378:metabolism / Q000648:ultrastructure*; D003208:Concanavalin A / Q000378:metabolism*; D008099:Liver / Q000648:ultrastructure; D051381:Rats; D011952:Receptors, Concanavalin A / Q000302:isolation & purification / Q000378:metabolism
D002843:Chromatin; D011952:Receptors, Concanavalin A; D003208:Concanavalin A