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Volume 270, Number 12, Issue of March 24, 1995 pp. 6768-6772
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
The Gap Junction Proteins -Connexin (Connexin-32) and -Connexin (Connexin-26) Can Form Heteromeric Hemichannels (*)

(Received for publication, October 3, 1994; and in revised form, December 14, 1994)

Kathrin A. Stauffer (§)

From the Medical Research Council Laboratory of Molecular Biology, Cambridge, United Kingdom

ABSTRACT

Two different types of gap junction proteins, beta(1)- and beta(2)-connexin, were expressed in insect cells, either singly or together, using infection with recombinant baculovirus. Membrane fractions enriched in gap junction proteins were isolated, and connexons (hemichannels) were solubilized with detergent. These solubilized connexons were then run out on a gel filtration column which was capable of partially separating the two homomeric connexons. It was found that connexons from cells co-infected with both types of baculovirus ran together on this column, whereas connexons from cells infected separately and mixed before solubilization did not, suggesting that in the co-infected cells the two types of connexin are assembled into heteromeric hemichannels.

FOOTNOTES

*
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked ``advertisement'' in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§
Present address: Dept. of Pathology, Cambridge University, Tennis Court Rd., Cambridge, UK. Tel.: 44-223-333-740; Fax: 44-223-333-732.

(^1)
The abbreviations used are: LC-14, monomyristoyl lysolecithin; PAGE, polyacrylamide gel electrophoresis.

(^2)
M. Cascio, N. M. Kumar, and N. B. Gilula, unpublished observations.

(^3)
N. C. Koenig, personal communication.

ACKNOWLEDGEMENTS

I acknowledge the generous gift of recombinant baculovirus, the cell line M12.13, and the antibody against the beta(2)-protein from Nalin Kumar, useful help with the culturing of M12.13 from Robert Safarik, and excellent secretarial assistance from Tim Green. Many helpful and stimulating discussions with Lukas Buehler, Bernie Gilula, Nalin Kumar, John Berriman, Tim Green, Nicola Koenig, and Olga Perisic were essential to this work, and I thank them all for their time and attention. Special thanks are due to Nigel Unwin for his unwavering support of this work.

©1995 by The American Society for Biochemistry and Molecular Biology, Inc.

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