Aplysia californica neurons R3-R14: primary structure of the myoactive histidine-rich basic peptide and peptide I.
The R3-R14 neurons of the marine mollusc Aplysia are neuroendocrine cells that express a gene encoding peptides I, II and histidine-rich basic peptide (HRBP), a myoactive peptide that excites Aplysia heart and enhances gut motility in vitro. Peptide II has been chemically characterized (35), but the complete primary structures of peptide I and HRBP have not been established by amino acid sequence analysis. HRBP, peptide I, and the prohormone (proHRBP) were therefore purified from acid extracts of Aplysia californica neural tissue using sequential gel filtration and reverse-phase high-performance liquid chromatography and chemically characterized. Amino acid sequence analysis demonstrated that HRBP was a 43-residue peptide whose sequence was: less than Glu-Val-Ala-Gln-Met-His-Val-Trp-Arg-Ala-Val-Asn-His-Asp-Arg-Asn-His-Gly- Thr-Gly - Ser-Gly-Arg-His-Gly-Arg-Phe-Leu-Ile-Arg-Asn-Arg-Tyr-Arg-Tyr-Gly-Gly-Gly- His-Leu - Ser-Asp-Ala-COOH. Compositional and sequence analyses of peptide I and proHRBP demonstrated that peptide I was a 26-residue peptide with the following sequence: NH2-Glu-Glu-Val-Phe-Asp-Asp-Thr-Asp-Val-Gly-Asp-Glu-Leu-Thr-Asn-Ala- Leu-Glu-Ser-Val-Leu-Thr-Asp-Phe-Lys-Asp-COOH. These results demonstrated that the pro-HRBP sequence predicted by nucleotide sequence analysis of a cDNA clone (24) was in fact synthesized in R3-R14 neurons. Hydrophilicity and hydrophobicity profiles of preproHRBP, combined with charge distribution profiles and predictive secondary structural analysis, showed that cleavage at dibasic sequences was strongly associated with peaks of hydrophilicity in alpha-helical regions of the preprohormone.
Nagle
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GT
Marine Biomedical Institute, University of Texas Medical Branch, Galveston 77550.
Knock
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SL
Painter
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SD
Blankenship
J E
JE
Fritz
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Kurosky
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A
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001048:Aplysia / Q000378:metabolism*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003488:Cyanogen Bromide; D006868:Hydrolysis; D008969:Molecular Sequence Data; D009474:Neurons / Q000032:analysis*; D009479:Neuropeptides / Q000032:analysis*; D011487:Protein Conformation; D011751:Pyroglutamyl-Peptidase I
D000596:Amino Acids; D009479:Neuropeptides; C061786:peptide I (Aplysia); C052709:histidine rich basic peptide (Aplysia); D011751:Pyroglutamyl-Peptidase I; D003488:Cyanogen Bromide