Two alloalbumins with identical electrophoretic mobility are produced by differently charged amino acid substitutions.
We describe the amino acid substitutions of albumins Sondrio and Paris 2, two slow moving variants of human serum albumin, which show an identical electrophoretic mobility on cellulose acetate at three different pH values. These variants have been found in several instances in a wide geographic area including Northern Italy and France. Both alloalbumins were isolated from the sera of heterozygous subjects. Isoelectric focusing analysis of CNBr fragments from the purified variants allowed us to localize the mutation of albumin Sondrio in fragment CNBr V (residues 330-446) and that of albumin Paris 2 in CNBr VII (residues 549-585). Sequential analysis of the variant CNBr VII established the molecular defect of albumin Paris 2 as 563 Asp----Asn. Fragments CNBr V from normal and Sondrio albumins were isolated on a preparative scale and subjected to tryptic and V8 proteinase digestion. Sequence determination of the abnormal tryptic and V8 peptides revealed that the variant arises from the substitution of glutamic acid 333 by lysine. Thus, a +1 change in the C-terminal region of the albumin molecule produces a variant with the same electrophoretic mobility as an alloalbumin with a +2 substitution in the central domain, suggesting a higher degree of exposure to the solvent of the C-terminal tailpiece. Both amino acid substitutions are consistent with a G----A transition in the first position of the corresponding codon in the structural gene.
Dipartimento di Biochimica, Università, Pavia, Italy.
L
L
Minchiotti
M
M
Galliano
M
M
Stoppini
G
G
Ferri
H
H
Crespeau
D
D
Rochu
F
F
Porta
D000595:Amino Acid Sequence; D001216:Asparagine / Q000737:chemistry; D001224:Aspartic Acid / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D014644:Genetic Variation; D005971:Glutamates / Q000737:chemistry; D018698:Glutamic Acid; D006801:Humans; D007525:Isoelectric Focusing; D008239:Lysine / Q000737:chemistry; D008969:Molecular Sequence Data; D009154:Mutation; D012709:Serum Albumin / Q000737:chemistry*
D005971:Glutamates; D012709:Serum Albumin; D001224:Aspartic Acid; D018698:Glutamic Acid; D001216:Asparagine; D008239:Lysine