TY - JOUR
T1 - The molecular chaperone calnexin associates with the vacuolar H+-ATPase from oat seedlings
AU - Li, Xuhang
AU - Su, Robert T.C.
AU - Hsu, Hei Ti
AU - Sze, Heven
PY - 1998
Y1 - 1998
N2 - Acidification of endomembrane compartments by the vacuolar-type H+-ATPase (V-ATPase) is central to many cellular processes in eukaryotes, including osmoregulation and protein sorting. The V-ATPase complex consists of a peripheral sector (V1) and a membrane integral sector (V(o)); however, it is unclear how the multimeric enzyme is assembled. A 64-kD polypeptide that had copurified with oat V-ATPase subunits has been identified as calnexin, an integral protein on the endoplasmic reticulum. To determine whether calnexin interacted physically with the V-ATPase, microsomal membranes were Triton X-100 solubilized, and the protein-protein interaction was analyzed by coimmunoprecipitation. Monoclonal antibodies against calnexin precipitated both calnexin and V-ATPase subunits, including A and B and those of 44, 42, 36, 16, and 13 kD. A monoclonal antibody against subunit A precipitated the entire V-ATPase complex as well as calnexin and BiP, an endoplasmic reticulum lumen chaperone. The results support our hypothesis that both calnexin and BiP act as molecular chaperones in the folding and assembly of newly synthesized V1V(o)-ATPases at the endoplasmic reticulum.
AB - Acidification of endomembrane compartments by the vacuolar-type H+-ATPase (V-ATPase) is central to many cellular processes in eukaryotes, including osmoregulation and protein sorting. The V-ATPase complex consists of a peripheral sector (V1) and a membrane integral sector (V(o)); however, it is unclear how the multimeric enzyme is assembled. A 64-kD polypeptide that had copurified with oat V-ATPase subunits has been identified as calnexin, an integral protein on the endoplasmic reticulum. To determine whether calnexin interacted physically with the V-ATPase, microsomal membranes were Triton X-100 solubilized, and the protein-protein interaction was analyzed by coimmunoprecipitation. Monoclonal antibodies against calnexin precipitated both calnexin and V-ATPase subunits, including A and B and those of 44, 42, 36, 16, and 13 kD. A monoclonal antibody against subunit A precipitated the entire V-ATPase complex as well as calnexin and BiP, an endoplasmic reticulum lumen chaperone. The results support our hypothesis that both calnexin and BiP act as molecular chaperones in the folding and assembly of newly synthesized V1V(o)-ATPases at the endoplasmic reticulum.
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U2 - 10.1105/tpc.10.1.119
DO - 10.1105/tpc.10.1.119
M3 - Article
C2 - 9477575
AN - SCOPUS:0031832284
SN - 1040-4651
VL - 10
SP - 119
EP - 130
JO - Plant Cell
JF - Plant Cell
IS - 1
ER -