The effect of glycation of CD59 on complement-mediated cytolysis.
Vascular proliferation is one of the major causes for morbidity and mortality in diabetes. However, the cellular and molecular mechanisms that link hyperglycemia to this complication remain unclear. In present study, we demonstrated by site-directed mutagenesis that mutated CD59 was more susceptible to glycation-inactivation for hyperglycemia. Mutated and wild-type CD59s were stably expressed in Chinese hamster ovary cells using the pALTER-MAX mammalian expression vector. Western blot, FACS and immunological fluorescence were conducted to confirm that CD59s were tethered to the plasma membrane. Compared to wild-type CD59, human CD59 mutants led to a significant increase in dye release assay. These results indicate that there may be some mutations of CD59 in diabetes population and the mutated CD59, which is more likely to be of glycation than the wild-type, may help to explain the distinct propensity of diabetes subjects to develop vascular proliferation complications.
Cheng
Ying
Y
Department of Immunology, Medical College of Qingdao University, Qingdao, Shandong, China.
Gao
Meihua
M
D000818:Animals; D000920:Antibody-Dependent Cell Cytotoxicity / Q000235:genetics*; D018957:CD59 Antigens / Q000235:genetics / Q000378:metabolism*; D016466:CHO Cells; D049109:Cell Proliferation; D003001:Cloning, Molecular; D003171:Complement Pathway, Classical; D003165:Complement System Proteins / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D048909:Diabetes Complications / Q000235:genetics* / Q000401:mortality; D003920:Diabetes Mellitus / Q000235:genetics; D004198:Disease Susceptibility; D015870:Gene Expression; D006031:Glycosylation; D006801:Humans; D006943:Hyperglycemia / Q000209:etiology; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D014652:Vascular Diseases / Q000235:genetics* / Q000401:mortality
D018957:CD59 Antigens; D003165:Complement System Proteins