Conserved neuron promoting activity in Drosophila and vertebrate laminin alpha 1

• Published in: The Journal of biological chemistry Vol. 271; no. 30; pp. 18074 - 18081
• Main Authors: Takagi, Y. (UCLA, Los Angeles, CA.), Nomizu, M, Gullberg, D, MacKrell, A.J, Keene, D.R, Yamada, Y, Fessler, J.H
• Format: Journal Article
• Language: English
• Published: 26-07-1996
• Subjects: CULTIVO DE CELULAS; CULTURE DE CELLULE; DIFERENCIACION CELULAR; DIFFERENCIATION CELLULAIRE; DROSOPHILA; NERF; NERVIOS; PEPTIDE; PEPTIDOS; PROTEINAS; PROTEINE; TEJIDOS ANIMALES; TISSU ANIMAL
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.271.30.18074

Drosophila S2 cells were transfected with constructs that code for two portions of the Drosophila laminin alpha chain. Construct rec alphaL coded for domains III, I/II, and G of laminin alpha. Construct rec alphaS 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 rec alphaL chain partly formed disulfide-linked heterotrimers with the endogenously produced beta and gamma laminin chains. Like normal Drosophila laminin, a substrate coating of either rec alphaL or rec alphaS supported neuron differentiation and neurite extension of primary Drosophila embryo cell cultures. However, at the same low concentrations, only Drosophila laminin-1, but neither rec alphaL nor rec alphaS supported myogenesis in these cultures. Previously, an overlapping set of dodecapeptides that covered a region of the murine laminin alpha 1 chain similar to rec alphaS 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 alpha homologues of the sex 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