The repair of the surface structure of animal cells.
Experiments were performed to determine if animal cells in culture possess specific mechanisms to repair surface molecules damaged by enzymes. The surface membranes of a primary cell culture, chick fibroblasts, a permanent hamster cell line, BHK21/C13, and its virally transformed counterpart, C13/B4 were damaged by exposure to trypsin or to neuraminidase. Following digestion with trypsin, the incorporation of radioactive amino acids or sugars into purified surface membrane of cells was monitored. No differences were noted in rates of incorporation when control and trypsin-damaged cells were compared. Neuraminidase damage to the surface of BHK21/C13 and C13/B4 cells was evidenced by altered gel filtration profiles of surface glycopeptides, i.e., delayed elution because of reduction in size. By labelling cells with 14C-L-fucose prior to neuraminidase treatment and following the incorporation of 3H-L-fucose into cell surface glycopeptides after neuraminidase digestion, we were able to monitor the synthesis and turnover of fucose-containing glycopeptides in the same cells. Gel filtration profiles indicated that little or no desialylated glycoproteins were resialylated (repaired) by specific replacement of sialic acid. Comparing neuraminidase-digested and control cells we observed no difference in rates of 3H-L-fucose incorporation or of 14C-L-fucose loss from these cells; nor did we find differences in the rate of incorporation of isotopic glucosamine into sialic acid. Neuraminidase treatment failed to alter the rate of cell growth or the pattern of isotopic incorporation into various cell surface components. These results support the suggestion that return of sialic acid (repair) was effected by turnover which serves as a non-specific repair mechanism to replace damaged cell surface molecules (Warren and Glick '68; Warren, '69).
D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D005944:Glucosamine / Q000378:metabolism; D006020:Glycopeptides / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism*; D007700:Kinetics; D007930:Leucine / Q000378:metabolism; D008970:Molecular Weight; D009439:Neuraminidase / Q000494:pharmacology*; D012794:Sialic Acids / Q000378:metabolism*; D014357:Trypsin / Q000494:pharmacology*
D006020:Glycopeptides; D012794:Sialic Acids; D009439:Neuraminidase; D014357:Trypsin; D007930:Leucine; D005944:Glucosamine