Avian riboflavinuria--XI. Immunological quantitation of cross-reacting liver proteins from normal, heterozygous, and mutant hens

Abstract

A genetic defect, avian riboflavinuria, was discovered in a strain of Single Comb White Leghorn chickens and has been attributed to the absence of functional riboflavin-binding protein (RBP). The ratio of functional RBP in blood, egg yolk, and egg white was 2 (RdRd) : 1 (Rdrd) : 0 (rdrd). The present study on non-riboflavin-binding, cross-reacting proteins (CRPs) from RdRd, Rdrd, and rdrd hens involved partial purification and immunochemical quantitation using antiserum to RBP. Immunoreactivities (microgram/g liver) of CRPs were found to be 2.6 (RdRd) : 1.3 (Rdrd) : 0.02 (rdrd). Reciprocal cross-reactions were observed with rabbit sera directed toward both RdRd CRP and RBP. Reaction of the CRPs with antiglycopeptide serum (specific for beta-linked galactose) showed that they were glycosylated. CRPs from RdRd and Rdrd hens had relative antigenicities similar to that of RBP (Krel approximately or equal to 1), while rdrd CRP had a significantly lower antigenicity (Krel = 0.003). The molecular weights of the CRPs as determined by SDS-polyacrylamide gel electrophoresis were as follows: RdRd = 31,6000, Rdrd = 30,900, and rdrd = 27,500. The molecular weight of egg yolk RBP was 34,7000 by the same method. The conclusion is drawn that the rd gene codes for a nonfunctional mutant protein, possibly an "altered precursor", that is different from RBP.