Intracellular association between UDP-glucose:glycoprotein glucosyltransferase and an incompletely folded variant of alpha1-antitrypsin.
Genetic variants of human alpha1-antitrypsin unable to fold into the native structural conformation are poorly secreted from hepatocytes. The molecular chaperone calnexin coimmunoprecipitates with secretion-incompetent variant null(Hong Kong) retained in stably transfected mouse hepatoma cells (Le, A., Steiner, J. L., Ferrell, G. A., Shaker, J. F., and Sifers, R. N. (1994) J. Biol. Chem. 269, 7514-7519). Mobilization of intracellular Ca2+ stores with metabolic poisons diminished interaction with calnexin and coincided with coimmuoprecipitation of a 150-kDa protein (p150). Mobilization of endoplasmic reticulum lumenal Ca2+ with thapsigargin, an inhibitor of the microsomal Ca2+ATPase, gave a similar result. Coimmunoprecipitation of p150 was specifically disrupted in response to incubation of the cell lysate with exogenous CaCl2. Finally, in ECL Western blotting, p150 was recognized by polyclonal antiserum against UDP-glucose:glycoprotein glucosyltransferase that likely functions in glycoprotein folding and quality control (Sousa, M. C., Ferrero-Garcia, M. A., and Parodi, A. J. (1992) Biochemistry 31, 97-105). The data are consistent with a model in which perturbation of endoplasmic reticulum Ca2+ results in a stable physical association between unfolded human alpha1-antitrypsin and UDP-glucose:glycoprotein glucosyltransferase.
Choudhury
P
P
Departments of Pathology and Cell Biology, Baylor College of Medicine, University of Texas, Houston, Texas 77030, USA.
Liu
Y
Y
Bick
R J
RJ
Sifers
R N
RN
D000818:Animals; D015153:Blotting, Western; D005964:Glucosyltransferases / Q000378:metabolism*; D006801:Humans; D051379:Mice; D017510:Protein Folding; D014407:Tumor Cells, Cultured; D014532:Uridine Diphosphate Glucose / Q000378:metabolism*; D000515:alpha 1-Antitrypsin / Q000737:chemistry / Q000378:metabolism*
D000515:alpha 1-Antitrypsin; D005964:Glucosyltransferases; D014532:Uridine Diphosphate Glucose