Purification, cloning and nucleotide sequence determination of cynomolgus monkey apolipoprotein C-II: comparison to the human sequence

• Published in: Molecular and cellular biochemistry 1989-10, Vol.90 (1), p.69-79
• Main Authors: Whitted, B E, Castle, C K, Polites, H G, Melchior, G W, Marotti, K R
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Apolipoprotein C-II; Apolipoproteins C - analysis; Apolipoproteins C - genetics; Apolipoproteins C - isolation & purification; Base Sequence; DNA - analysis; DNA - isolation & purification; genes; Humans; Macaca fascicularis; Molecular Sequence Data; RNA - analysis; RNA - isolation & purification; Sequence Homology, Nucleic Acid
• ISSN: 0300-8177

We have purified apolipoprotein C-II (apo C-II) from cynomolgus monkey plasma, prepared antibody against it and used the antibody to isolate a cDNA containing the complete coding sequence for cynomolgus monkey apo C-II. Sequence analysis indicated that the monkey apo C-II cDNA was 200 bp longer than the human and the difference in size was all in the 5 degrees untranslated region of mRNA. This was confirmed by Northern analysis of human and monkey RNA. There was an open reading frame in the monkey apo C-II cDNA sequence encoding a preprotein of 101 amino acids - identical in size to the human protein. The carboxyl terminal 44 amino acids of the protein were 100% homologous to the human apo C-II amino acid sequence indicating evolutionary conservation of both structure and function. However, the amino terminal 35 amino acids of the protein were only 75% homologous and the amino terminal 19 amino acids were only 58% homologous to the human sequence. The amino acid sequence derived from the nucleotide sequence predicts a more basic protein than the human apo C-II and this is confirmed by isoelectric focusing and immunoblotting.