Molecular and functional characterization of amylin, a peptide associated with type 2 diabetes mellitus.
The 37-amino acid peptide called amylin is a major component of the islet amyloid deposited in the pancreases of persons with type 2 diabetes mellitus. We report the isolation of a partial cDNA clone and a phage lambda genomic clone of the coding region of the amylin gene. The DNA sequence encodes a protein sequence identical to that of amylin isolated from the amyloid found in the diabetic pancreas and shows that amylin is likely to be synthesized as a precursor peptide, now named proamylin. We have demonstrated that the amylin gene is present on chromosome 12 and that it is probably transcribed in the islets of Langerhans. The sequences of the genes for amylin and the calcitonin gene-related peptides (CGRPs) show strong similarity, especially over their 5' coding regions, where both peptides have a conserved intramolecular disulfide bridge, and also over their 3' coding regions, where the presence of a glycine codon strongly suggests that the carboxyl-terminal residue of amylin, like that of CGRP, is amidated. To examine the functional relevance of these posttranslational modifications, the biological activity of amylin synthesized with or without the disulfide bridge and/or amidation was measured. It was found that both features are necessary for full biological activity, thereby confirming the functional importance of those regions of the molecule whose sequences are conserved at both protein and genetic levels.
Roberts
A N
AN
Department of Biochemistry, University of Oxford, United Kingdom.
Leighton
B
B
Todd
J A
JA
Cockburn
D
D
Schofield
P N
PN
Sutton
R
R
Holt
S
S
Boyd
Y
Y
Day
A J
AJ
Foot
E A
EA
D000595:Amino Acid Sequence; D000682:Amyloid / Q000235:genetics* / Q000302:isolation & purification; D001483:Base Sequence; D015740:Calcitonin Gene-Related Peptide / Q000235:genetics; D002881:Chromosomes, Human, Pair 12; D003001:Cloning, Molecular; D004247:DNA / Q000096:biosynthesis; D003924:Diabetes Mellitus, Type 2 / Q000235:genetics* / Q000378:metabolism; D015894:Genome, Human; D006801:Humans; D058228:Islet Amyloid Polypeptide; D007515:Islets of Langerhans / Q000032:analysis / Q000378:metabolism*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D012333:RNA, Messenger / Q000235:genetics; D012016:Reference Values; D012689:Sequence Homology, Nucleic Acid
D000682:Amyloid; D058228:Islet Amyloid Polypeptide; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D004247:DNA; D015740:Calcitonin Gene-Related Peptide