Characterization of a putative molluscan insulin-related peptide receptor.
In the pond snail Lymnaea stagnalis (Ls), growth and associated processes are likely to be controlled by a family of molluscan insulin-related peptides (MIP). Here we report on the cloning of a cDNA encoding a putative receptor for these MIP. This cDNA was isolated from Ls via PCR with degenerate oligodeoxynucleotides corresponding to conserved parts of the tyrosine kinase domain of the human insulin receptor and its Drosophila homologue. Many of the typical insulin-receptor features, including a cysteine-rich domain, a single transmembrane domain and a tyrosine-kinase domain are conserved in the predicted, 1607-amino acid (aa) protein. Comparison of the aa sequence of the molluscan receptor to other insulin-receptor sequences revealed strong variations in the percentage of sequence identity for the different domains, ranging from 70% sequence identity in the tyrosine-kinase domain to virtually no sequence identity in the C-terminal sequence. Striking differences are the absence of a clear tetrabasic cleavage site, and the extremely long C-terminus of 308 aa that contains seven Tyr residues. Southern blot analyses at varying stringencies, extensive screening of cDNA- and genomic libraries, and PCR experiments indicate the presence of a single putative MIP receptor. This suggests that the four different MIP may exert their functional role in Ls by binding to the same receptor.
Roovers
E
E
Department of Biochemistry and Molecular Biology, Vrije Universiteit, Amsterdam, The Netherlands.
Vincent
M E
ME
van Kesteren
E
E
Geraerts
W P
WP
Planta
R J
RJ
Vreugdenhil
E
E
van Heerikhuizen
H
H
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000235:genetics; D005796:Genes; D008195:Lymnaea / Q000235:genetics*; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000378:metabolism*; D011972:Receptor, Insulin / Q000235:genetics*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction
D017931:DNA Primers; D018076:DNA, Complementary; D009479:Neuropeptides; C063879:insulin-related neuropeptide; D011972:Receptor, Insulin