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Volume 271, Number 30, Issue of July 26, 1996 pp. 18074-18081
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

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Conserved Neuron Promoting Activity in Drosophila and Vertebrate Laminin 1

(Received for publication, December 13, 1995, and in revised form, April 29, 1996)

Yasumitsu Takagi , Motoyoshi Nomizu ^¶ , Donald Gullberg , Albert J. MacKrell , Douglas R. Keene , Yoshihiko Yamada ^¶ and John H. Fessler

From the  Molecular Biology Institute and Biology Department, UCLA, Los Angeles, California 90095-1570,  The Shriners Hospital for Crippled Children, Portland, Oregon 97201, and the ^¶ Laboratory of Developmental Biology, NIDR, National Institutes of Health, Bethesda, Maryland 20892

Drosophila S2 cells were transfected with constructs that code for two portions of the Drosophila laminin  chain. Construct recL coded for domains III, I/II, and G of laminin . Construct recS coded for only the COOH-most 12% of the I/II domain and the G domain. The corresponding polypeptides were isolated and characterized from the culture media. The recL chain partly formed disulfide-linked heterotrimers with the endogenously produced  and  laminin chains. Like normal Drosophila laminin, a substrate coating of either recL or recS supported neuron differentiation and neurite extension of primary Drosophila embryo cell cultures. However, at the same low concentrations, only Drosophila laminin-1, but neither recL nor recS supported myogenesis in these cultures. Previously, an overlapping set of dodecapeptides that covered a region of the murine laminin 1 chain similar to recS had been synthesized and tested for cell culture support properties (Nomizu, M., Kim, W. H., Yamamura, K., Utani, A., Otaka, A., Roller, P. P., Kleinman, H. K., and Yamada, Y. (1995) J. Biol. Chem. 270, 20583-20590). The Drosophila laminin  homologues of the six most active vertebrate dodecapeptides were now synthesized and tested as substrates for differentiation of primary Drosophila embryo cells. Peptides that contained either the Drosophila sequence SIKVGV or the murine homologue, SIKVAV, provided support for neurite extension.

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