Aplysia temptin - The 'glue' in the water-borne attractin pheromone complex

Scott F. Cummins, Fang Xie, Melissa R. De Vries, Suresh P. Annangudi, Milind Misra, Bernard M. Degnan, Jonathan V. Sweedler, Gregg T. Nagle, Catherine H. Schein

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Temptin, a component of the complex of water-borne protein pheromones that stimulate attraction and mating behavior in the marine mollusk Aplysia, has sequence homology to the epidermal growth factor (EGF)-like domains of higher organisms that mediate protein-cell surface contact during fertilization and blood coagulation. In this work, recombinant temptin for structural and functional studies was produced in Escherichia coli using a cold shock promoter and purified by RP-HPLC. CD spectra confirmed a predominantly β-sheet structure. Two disulfide bonds were determined via limited proteolysis and MS. One internal disulfide (Cys57-Cys77) was predicted from initial alignments with class I EGF-like domains; the second, between Cys18 and Cys103, could protect temptin against proteolysis in seawater and stabilize its interacting surface. A three-dimensional model of temptin was prepared with our MPACK suite, based on the Ca2+-binding, EGF-like domain of the extracellular matrix protein fibrillin. Two temptin residues, Trp52 and Trp79, which align with cysteine residues conserved in fibrillins, lie adjacent to and could stabilize the disulfide bonds and a proposed metal-binding loop. The water-borne pheromone attractin in egg cordon eluates is complexed with other proteins. Docking results with our model and the NMR structure of attractin suggest that one face of temptin interacts with the pheromone, perhaps controlling its access to the cellular receptors. Gel shifts confirmed that temptin complexes with wild-type attractin. These results indicate that temptin, analogous to the role of fibrillin in controlling transforming growth factor-β concentration, modulates pheromone signaling by direct binding to attractin.

Original languageEnglish (US)
Pages (from-to)5425-5437
Number of pages13
JournalFEBS Journal
Volume274
Issue number20
DOIs
StatePublished - Oct 1 2007

Fingerprint

Aplysia
Glues
Pheromones
Adhesives
Epidermal Growth Factor
Disulfides
Proteolysis
Water
Mollusca
Extracellular Matrix Proteins
Seawater
Transforming Growth Factors
Blood Coagulation
Sequence Homology
Coagulation
Fertilization
Escherichia coli
Ovum
Cysteine
Shock

Keywords

  • Enticin
  • Epidermal growth factor-like domains
  • Fibrillin
  • Marfan's syndrome
  • Signaling network

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Cummins, S. F., Xie, F., De Vries, M. R., Annangudi, S. P., Misra, M., Degnan, B. M., ... Schein, C. H. (2007). Aplysia temptin - The 'glue' in the water-borne attractin pheromone complex. FEBS Journal, 274(20), 5425-5437. https://doi.org/10.1111/j.1742-4658.2007.06070.x

Aplysia temptin - The 'glue' in the water-borne attractin pheromone complex. / Cummins, Scott F.; Xie, Fang; De Vries, Melissa R.; Annangudi, Suresh P.; Misra, Milind; Degnan, Bernard M.; Sweedler, Jonathan V.; Nagle, Gregg T.; Schein, Catherine H.

In: FEBS Journal, Vol. 274, No. 20, 01.10.2007, p. 5425-5437.

Research output: Contribution to journalArticle

Cummins, SF, Xie, F, De Vries, MR, Annangudi, SP, Misra, M, Degnan, BM, Sweedler, JV, Nagle, GT & Schein, CH 2007, 'Aplysia temptin - The 'glue' in the water-borne attractin pheromone complex', FEBS Journal, vol. 274, no. 20, pp. 5425-5437. https://doi.org/10.1111/j.1742-4658.2007.06070.x
Cummins SF, Xie F, De Vries MR, Annangudi SP, Misra M, Degnan BM et al. Aplysia temptin - The 'glue' in the water-borne attractin pheromone complex. FEBS Journal. 2007 Oct 1;274(20):5425-5437. https://doi.org/10.1111/j.1742-4658.2007.06070.x
Cummins, Scott F. ; Xie, Fang ; De Vries, Melissa R. ; Annangudi, Suresh P. ; Misra, Milind ; Degnan, Bernard M. ; Sweedler, Jonathan V. ; Nagle, Gregg T. ; Schein, Catherine H. / Aplysia temptin - The 'glue' in the water-borne attractin pheromone complex. In: FEBS Journal. 2007 ; Vol. 274, No. 20. pp. 5425-5437.
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AU - Annangudi, Suresh P.

AU - Misra, Milind

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