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Molecular cloning of four cDNAs encoding prepro-crustacean hyperglycemic hormone (CHH) from the eyestalk of the red rock crab Cancer productus: identification of two genetically encoded CHH isoforms and two putative post-translationally derived CHH variants.
- Source :
-
General and comparative endocrinology [Gen Comp Endocrinol] 2008 Feb 01; Vol. 155 (3), pp. 517-25. Date of Electronic Publication: 2007 Sep 19. - Publication Year :
- 2008
-
Abstract
- Recently, we demonstrated that the four known sinus gland (SG) isoforms of Cancer productus crustacean hyperglycemic hormone precursor-related peptide (Capr-CPRP I-IV) are differentially distributed in conserved patterns among individual crabs. This finding strongly supported the presence of multiple prepro-crustacean hyperglycemic hormone (chh) transcripts in each crab, as well as the translation and processing of the encoded prepro-hormones. Whether these transcripts contained common or distinct isoforms of CHH remained unknown. To address this question, molecular analyses of the C. productus eyestalk prepro-chhs were undertaken. Using a PCR-based cloning strategy, four prepro-chh cDNAs were characterized: one encoding CPRP I, one encoding CPRP III (found to possess Ile(26) rather than Leu(26) as reported previously), and two encoding CPRP II. No cDNA encoding CPRP IV was identified. The deduced CHH present in the prepro-hormones containing CPRP I and III were identical (Capr-CHH I) and differed from that (Capr-CHH II) present in the two prepro-hormones containing Capr-CPRP II at a single residue, a Thr(5) for Ser(5) substitution. As both CHH isoforms possess Glu at position 1, a cyclization of this residue to pyroglutamine is likely as the peptides mature, as has been seen for the CHHs of other brachyuran species. Likewise, homology to other CHHs suggests all C. productus isoforms are C-terminally amidated. These post-translational modifications would result in four SG isoforms of CHH: Capr-CHH I, Capr-pyro-CHH I, Capr-CHH II, and Capr-pyro-CHH II. Southern blotting supported the hypothesis that at least three prepro-chh transcripts are present in each crab, while dual in situ hybridization-immunohistochemistry localized the transcripts to previously mapped CHH immunopositive somata in the X-organ, the major source of innervation to the SG.
- Subjects :
- Amino Acid Sequence
Animals
Base Sequence
Cloning, Molecular
DNA, Complementary isolation & purification
Eye chemistry
Eye metabolism
Molecular Sequence Data
Peptide Hormones genetics
Peptide Hormones metabolism
Protein Isoforms genetics
Protein Isoforms metabolism
Sequence Homology, Amino Acid
Brachyura genetics
Endocrine Glands metabolism
Nerve Tissue Proteins genetics
Nerve Tissue Proteins metabolism
Protein Precursors genetics
Protein Precursors metabolism
Protein Processing, Post-Translational
Subjects
Details
- Language :
- English
- ISSN :
- 0016-6480
- Volume :
- 155
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- General and comparative endocrinology
- Publication Type :
- Academic Journal
- Accession number :
- 17961562
- Full Text :
- https://doi.org/10.1016/j.ygcen.2007.09.001