Conserved neuron promoting activity in Drosophila and vertebrate laminin alpha1.
Drosophila S2 cells were transfected with constructs that code for two portions of the Drosophila laminin alpha chain. Construct recalphaL coded for domains III, I/II, and G of laminin alpha. Construct recalphaS 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 recalphaL chain partly formed disulfide-linked heterotrimers with the endogenously produced beta and gamma laminin chains. Like normal Drosophila laminin, a substrate coating of either recalphaL or recalphaS supported neuron differentiation and neurite extension of primary Drosophila embryo cell cultures. However, at the same low concentrations, only Drosophila laminin-1, but neither recalphaL nor recalphaS supported myogenesis in these cultures. Previously, an overlapping set of dodecapeptides that covered a region of the murine laminin alpha1 chain similar to recalphaS 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 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.
Takagi
Y
Y
Molecular Biology Institute and Biology Department, UCLA, Los Angeles, California 90095-1570, USA.
Nomizu
M
M
Gullberg
D
D
MacKrell
A J
AJ
Keene
D R
DR
Yamada
Y
Y
Fessler
J H
JH
D000595:Amino Acid Sequence; D000818:Animals; D001681:Biological Assay; D002448:Cell Adhesion / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D017124:Conserved Sequence; D004305:Dose-Response Relationship, Drug; D004330:Drosophila; D004622:Embryo, Mammalian / Q000166:cytology; D004625:Embryo, Nonmammalian; D004848:Epithelium / Q000187:drug effects; D007797:Laminin / Q000235:genetics / Q000494:pharmacology* / Q000648:ultrastructure; D051379:Mice; D008969:Molecular Sequence Data; D009132:Muscles / Q000187:drug effects; D009474:Neurons / Q000187:drug effects*; D009842:Oligopeptides / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013329:Structure-Activity Relationship
D007797:Laminin; D009842:Oligopeptides; D011994:Recombinant Proteins