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ELECTROPHORETIC ANALYSIS OF MELANOGENIC PROTEINS FROM NORMAL AND MUTANT CHICK MELANOCYTES
Abstract
Two-dimensional electrophoretic studies were done on three albino mutants of the fowl, Gallus gallus, and compared to the standard genotype. The mutants were: recessive white (c), autosomal albinism (a) and sex-linked albinism (s('al)). Two of these mutants, recessive white and autosomal albinism, were dopa-oxidase negative and one, sex-linked albinism, was dopa-oxidase positive. Allelism tests between the two dopa-oxidase negative mutants showed the two genes to be alleles. A change in the genetic symbol from a to c('a) is proposed. Chicken tyrosinase activity was analyzed and the enzyme isolated. Tyrosinase activity of sex-linked albinism was found to be greater than twice that of the standard type, but absent in both c and c('a). Its location on a denaturing two-dimensional gel was determined. The enzyme was found to be a series of isozymes with a pI range of 4 to 4.5 and a molecular weight of 72,000 daltons. Cell cultures of neural crest cells of the standard type and of the mutants c, c('a), and s('al) were done. 4-(beta)-phorbol-12-(beta)-myristate-13-(alpha)-acetate (TPA) was used as a mitogen. No pigment production was observed in the standard type in the presence of TPA, but pigment production was observed when TPA was removed. No pigment was found in any of the mutant genotypes while in TPA, or when TPA was removed, but the mutant cells changed morphologically to the differentiated state upon removal of TPA. Two-dimensional electrophoresis was done on the standard type and the mutants utilizing neural crest cultures labeled with ('3)H-leucine and the proteins detected using fluorography. All four genotypes had a row of spots that co-migrated to the same location as isolated tyrosinase. A comparison of the protein patterns between cells grown in TPA and when TPA was removed was made. A number of proteins that were TPA sensitive, including tyrosinase, were found and are proposed to be putative melanogenic proteins.
Subject Area
Genetics
Recommended Citation
OETTING, WILLIAM STARR, "ELECTROPHORETIC ANALYSIS OF MELANOGENIC PROTEINS FROM NORMAL AND MUTANT CHICK MELANOCYTES" (1983). ETD collection for University of Nebraska-Lincoln. AAI8314910.
https://digitalcommons.unl.edu/dissertations/AAI8314910