Nature 277, 403 - 405 (01 February 1979); doi:10.1038/277403a0

Isolation of the human placenta receptor for epidermal growth factor-urogastrone

R. ALAN HOCK, EBBA NEXØ & MORLEY D. HOLLENBERG

Howard Hughes Medical Institute, Division of Clinical Pharmacology, Department of Medicine and Department of Pharmacology and Experimental Therapeutics, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205

EPIDERMAL growth factor-urogastrone (EGF-URO), a singlechain polypeptide of molecular weight 6,000, found both in the mouse¹⁻³ and in man⁴⁻⁷, is both a potent stimulant of cell proliferation and an inhibitor of gastric acid secretion. Specific membrane receptors for EGF-URO can be detected in various tissues including human placenta⁸⁻¹¹. Although the mouse and human polypeptide sequences differ at 16 positions, they can share the same receptor site in human tissues^3,12. There has been considerable recent interest in EGF-URO and its receptor, not only because of the potent mitogenic and acid-inhibitory actions of the polypeptide, but also because of the association of viral¹³, chemical¹⁴ and spontaneous¹⁵ cell transformation with a specific reduction in cell receptors for EGF-URO. It is also believed that EGF-URO and its receptor may be involved in the development, maintenance, and differentiation of tissues as diverse as the lens¹⁶, the palate¹⁷ and the keratin layer of skin¹⁸. In experiments aimed at the isolation of the membrane receptor for EGF-URO, we have observed that, unlike the receptor for insulin¹⁹, but like the muscarinic receptor for acetylcholine²⁰, the EGF-URO receptor loses its ligand recognition property following solubilisation with detergents. We therefore developed an affinity-labelling technique using glutaraldehyde, that can covalently crosslink the receptor with ¹²⁵I-labelled EGF-URO before solubilisation²¹. Independently, Das and coworkers have developed a photoaffinity-labelling technique that can also be used to radiolabel the EGF-URO receptor before solubilisation²². Here we report the success of both our previously described glutaraldehyde crosslinking technique²¹ (using ¹²⁵I-EGF-URO for receptor labelling) and of a newly developed photoaffinity-labelling technique (using a photolabile ¹²⁵I-EGF-URO crosslinking reagent) to identify the EGF-URO receptor in human placenta membranes and to effect the isolation of the affinity-labelled receptor by affinity chromatography using lectin and murine EGF-URO antibody−Agarose columns.

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