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Isolation of an egg-laying hormone-binding protein from the gonad of Aplysia californica and its localization in oocytes

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Abstract

A protein solubilized from a membrane preparation of the gonad of Aplysia californica has been isolated by affinity chromatography, using bag cell egg-laying hormone (ELH) as the bound ligand, and partially purified and characterized by gel electrophoresis. The protein has an apparent molecular weight of 52 kDa and consists of two disulfide-linked subunits of about 30 kDa each. The protein is glycosylated and has an acidic pI. Approximately 10–15 μg of this protein can be isolated from a single ovotestis, representing less than 1% of the total protein in the gonad; but the protein could not be detected in buccal mass or body wall, tissues which do not have apparent response to ELH.

Antibodies generated against this ELH-binding protein (ELHBP) were used to localize sites in the ovotestis which might contain this molecule and thus represent targets for egg-laying hormone. Immunocytochemical results indicate that the oocytes are a rich source of this protein, since their cytoplasm was the only detectable site of immunoreactivity.

Whether this binding protein represents an egg-laying hormone receptor is uncertain, but its prevalence in oocytes suggests that ELH plays a signaling role on these gametes.

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Abbreviations

ConA:

convalin A

DAB:

diaminobenzidine

ELH:

egg-laying hormone

ELHBP:

ELH-binding protein

IEF:

isoelectric focusing

IGFR:

insulin-like growth factor

IgG:

immunoglobulin

NGS:

normal goat serum

PAGE:

polyacrylamide gel electrophoresis

SDS:

sodium dodecylsulfate

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Author notes

  1. J. V. A. Choate

    Present address: Department of Physiology, Oregon Health Science University, 97201, Portland, OR, USA

  2. T. E. Kruger

    Present address: Department of Ophthalmology, Louisiana State University Medical Center, 71130, Shreveport, LA, USA

  3. M. -A. Micci

    Present address: , Via A. Poerio, 45, I-00152, Rome, Italy

Authors and Affiliations

  1. Marine Biomedical Institute, University of Texas Medical Branch, 77555-0843, Galveston, TX, USA

    J. V. A. Choate, T. E. Kruger, M. -A. Micci & J. E. Blankenship

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Choate, J.V.A., Kruger, T.E., Micci, M.A. et al. Isolation of an egg-laying hormone-binding protein from the gonad of Aplysia californica and its localization in oocytes. J Comp Physiol A 173, 475–483 (1993). https://doi.org/10.1007/BF00193520

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