Please use this identifier to cite or link to this item: https://doi.org/10.1016/S1357-2725(98)00057-0
Title: Functional role of glycosylation on the recombinant CD38/ADP-ribosyl cyclase in CHO cells
Authors: Chidambaram, N.
Chang, C.F. 
Keywords: ADP-ribosyl cyclase
CD38
CHO cells
deglycosylation
enzyme kinetics
Issue Date: 1998
Source: Chidambaram, N.,Chang, C.F. (1998). Functional role of glycosylation on the recombinant CD38/ADP-ribosyl cyclase in CHO cells. International Journal of Biochemistry and Cell Biology 30 (9) : 1011-1018. ScholarBank@NUS Repository. https://doi.org/10.1016/S1357-2725(98)00057-0
Abstract: Current evidence suggests that CD38, a lymphocyte differentiation antigen, has been found to be functionally indistinguishable from the native form when it was expressed as a soluble and non-glycosylated protein in yeast. Studies were conducted to evaluate the functional role of glycosylation on the membrane-bound CD38 in mammalian cells. The stable transformants of CHO cells were established with pXJ41-CD38 construct and the recombinant CD38 was detected at the surface of CHO cells using a polyclonal antibody to rat CD38 by immunocytochemistry. The recombinant protein displaying ADP-ribosyl cyclase activity was purified from CHO cells, which appeared as 42 and 46 kDa bands on immunoblot under non-reducing and reducing conditions, respectively. The recombinant CD38 was then subjected to deglycosylation with N-glycosidase F that resulted in a 33 kDa band on immunoblot under reducing condition. Further the partial deglycosylation of the recombinant CD38, performed at various time intervals, resulted in a series of bands (33-46 kDa) on immunoblot. Kinetic analysis indicated that deglycosylation of the recombinant CD38 showed a considerable decrease in V(max) and an increase in K(m) of ADP-ribosyl cyclase activity. These observations clearly suggest that glycosylation plays an important role to maintain the enzymatic activity and substrate affinity of CD38/ADP-ribosyl cyclase for mediating the signalling events in mammalian cells. Copyright (C) 1998 Elsevier Science Ltd.
Source Title: International Journal of Biochemistry and Cell Biology
URI: http://scholarbank.nus.edu.sg/handle/10635/38180
ISSN: 13572725
DOI: 10.1016/S1357-2725(98)00057-0
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