Laminin carbohydrates are implicated in cell signaling.
We have examined how laminin carbohydrates participate in cellular responses and have focused upon cell spreading and neurite outgrowth. Our earlier studies showed that unglycosylated laminin fully supported cell adhesion but did not promote subsequent spreading of mouse melanoma cells or neurite outgrowth of rat pheochromocytoma cells (Dean et al. (1990): J Biol Chem 265:12553-12562). In the present experiments, we determined whether those cellular responses could be restored to adherent cells. When a mixture of unglycosylated and glycosylated laminins was used as a substratum for mouse melanoma cells, some cells began to spread when 30% glycosylated laminin was present. At least 65% glycosylated laminin was required to elicit a maximal spreading response by the majority of the cells. In separate experiments, we found that cell spreading was fully restored by a pronase digest of glycosylated laminin; a similar digest of unglycosylated laminin had no effect. These results indicate that laminin carbohydrates, rather than polypeptide sequences, were responsible for cell spreading. We also conclude that substrate attachment of the carbohydrate moieties was not essential. In other experiments, laminins containing immature oligosaccharides were produced using two glycosylation pathway inhibitors, swainsonine or castanospermine. When such laminins were used to study cell spreading or neurite outgrowth, laminin containing immature oligosaccharides was as effective as laminin which contains fully processed oligosaccharides. In contrast, laminin with partially processed oligosaccharides had incomplete activity. These composite reconstitution experiments show that laminin carbohydrates provide essential information to responsive cells, enabling them to progress from an adherent state to a spread form or to extend neurite processes.
Chandrasekaran
S
S
Department of BioStructure & Function, School of Dental Medicine, University of Connecticut Health Center, Farmington 06030.
Dean
J W
JW
Giniger
M S
MS
Tanzer
M L
ML
D000818:Animals; D001369:Axons / Q000502:physiology; D002448:Cell Adhesion / Q000187:drug effects* / Q000502:physiology; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D007212:Indolizines / Q000494:pharmacology; D007797:Laminin / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D008546:Melanoma, Experimental; D051379:Mice; D009474:Neurons / Q000166:cytology; D009844:Oligosaccharides / Q000494:pharmacology*; D010673:Pheochromocytoma; D011402:Pronase / Q000378:metabolism; D051381:Rats; D017026:Swainsonine / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D054836:Indolizidines
D007212:Indolizines; D007797:Laminin; D009844:Oligosaccharides; D011402:Pronase; D017026:Swainsonine; C037806:castanospermine; D054836:Indolizidines