Characterization of Aplysia Alb-1, a candidate water-borne protein pheromone released during egg laying

Scott F. Cummins, Bernard M. Degnan, Gregg T. Nagle

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Pheromones are known to be important to the innate behavior of marine animals. Attraction in Aplysia involves the long-distance water-borne protein pheromones attractin, enticin, temptin and seductin, which are released from the albumen gland during egg laying. Other pheromones are predicted to act in concert with these pheromones, but their identities are unknown. To identify additional pheromone candidates, we employed differential library screening of an albumen gland cDNA library, RT-PCR, recombinant protein expression, rhinophore contraction bioassays and immunocytochemistry. Alb-1 is expressed in the Aplysia californica albumen gland and encodes a novel protein that does not share significant sequence identity with any proteins in the database. RT-PCR analysis detected Alb-1 transcripts in the albumen gland, exocrine atrial gland and ovotestis. The Alb-1 precursor has a signal peptide sequence followed by a predicted 101-residue protein sequence containing eight cysteine residues. Recombinant protein expression, RP-HPLC, microsequence analysis and MALDI mass spectrometry analyses demonstrated that mature recombinant Alb-1 was processed at a paired basic residue site to generate an N-terminal and C-terminal protein fragment; this was consistent with immunoblot observations on purified albumen gland extracts. In rhinophore contraction (twitch) bioassays, the recombinant N-terminal protein induced rhinophore contractions whereas the C-terminal protein did not. An antibody generated to the N-terminal protein was used for immunocytochemical and immunoblot analyses and demonstrated that this protein is present in albumen gland secretory cells, egg cordons and egg eluates. Overall, the data suggest that Alb-1 may be processed in the albumen gland and that the Alb-11-56 protein released during egg laying may serve a pheromonal function in concert with attractin, enticin, temptin and seductin.

Original languageEnglish (US)
Pages (from-to)152-161
Number of pages10
JournalPeptides
Volume29
Issue number2
DOIs
StatePublished - Feb 1 2008

Fingerprint

Aplysia
Pheromones
Ovum
Water
Proteins
Protein C
Recombinant Proteins
Biological Assay
Bioassay
Exocrine Glands
Instinct
Polymerase Chain Reaction
Protein Databases
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Protein Sorting Signals
Gene Library
Cysteine
Mass Spectrometry
Immunohistochemistry
High Pressure Liquid Chromatography

Keywords

  • Alb-1
  • Aplysia
  • Attractin
  • Mollusk
  • Protein pheromone

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Endocrinology
  • Cellular and Molecular Neuroscience

Cite this

Characterization of Aplysia Alb-1, a candidate water-borne protein pheromone released during egg laying. / Cummins, Scott F.; Degnan, Bernard M.; Nagle, Gregg T.

In: Peptides, Vol. 29, No. 2, 01.02.2008, p. 152-161.

Research output: Contribution to journalArticle

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