cDNA and protein sequence of polymorphic macaque albumins that differ in bilirubin binding.
The rhesus monkey, Macaca mulatta, exhibits a geographically restricted polymorphism of serum albumins Mac A and Mac B that is recognized by electrophoresis and is associated with a difference in bilirubin-binding parameters. To identify the basis of the polymorphism, the cDNA and protein sequences of serum albumin from M. mulatta were determined. Screening of a lambda gt11 rhesus liver cDNA library yielded a 1988-bp cDNA sequence that encodes the complete amino acid sequence of mature albumin, the entire propeptide, and part of the prepropeptide. Isoelectric focusing and amino-terminal protein sequencing of CNBr fragments of albumin from A/A and B/B homozygotes were performed, and the structural difference was localized to a CNBr fragment (MCB3) spanning residues 124-264. Sequence analysis of lysyl endopeptidase peptides of MCB3 established that Mac A albumin has a glutamine residue at position 188 while the Mac B albumin has a glutamic residue at the same position. PCR amplification, subcloning, and DNA sequence analysis of clones from A/A and B/B homozygotes confirmed the protein sequence data and the codon difference of CAA versus GAA, respectively. Comparison of macaque and human serum albumin shows a 93.5% identity at the amino acid level. In human serum albumin, Glu188 is located close to the IIA binding pocket for ligands, probably including bilirubin. Derivatives of coumarin compete more efficiently with bilirubin for binding sites on the Mac A albumin than on the Mac B albumin. In regions where coumarin-containing plants are important food resources, Mac B albumin may confer a selective advantage because bilirubin is less readily displaced from it.
Watkins
S
S
Department of Biology, Indiana University, Bloomington 47405.
Sakamoto
Y
Y
Madison
J
J
Davis
E
E
Smith
D G
DG
Dwulet
J
J
Putnam
F W
FW
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001663:Bilirubin / Q000097:blood*; D001665:Binding Sites; D005075:Biological Evolution; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D005843:Geography; D006801:Humans; D008251:Macaca / Q000235:genetics; D008253:Macaca mulatta / Q000235:genetics*; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000379:methods; D011110:Polymorphism, Genetic; D017386:Sequence Homology, Amino Acid; D012709:Serum Albumin / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
D009838:Oligodeoxyribonucleotides; D012709:Serum Albumin; D004247:DNA; D001663:Bilirubin