TY - JOUR
T1 - Reconstitution of recombinant 40-kDa subunit of the clathrin-coated vesicle H+-ATPase
AU - Peng, Sheng Bin
AU - Stone, Dennis K.
AU - Xie, Xiao Song
N1 - Copyright:
Copyright 2004 Elsevier B.V., All rights reserved.
PY - 1993/11/5
Y1 - 1993/11/5
N2 - We have proposed a model of the ATP hydrolytic sector of the clathrin-coated vesicle H+-ATPase wherein significant catalysis requires four subunits of molecular masses of 70, 58, 40, and 33 kDa (Xie, X.-S., and Stone, D. K. (1988) J. Biol. Chem, 263, 9859-9867). We have cloned and expressed the 40-kDa component in Escherichia coli and have purified the recombinant protein to homogeneity. This subunit lacks ATP hydrolytic capacity, but when reconstituted to a 40 kDa-depleted hydrolytic sector, there is a greater than 20-fold increase in calcium-activated, N-ethylmaleimide-sensitive ATP hydrolysis, indicating that this subunit is required for vacuolar-type proton pump function.
AB - We have proposed a model of the ATP hydrolytic sector of the clathrin-coated vesicle H+-ATPase wherein significant catalysis requires four subunits of molecular masses of 70, 58, 40, and 33 kDa (Xie, X.-S., and Stone, D. K. (1988) J. Biol. Chem, 263, 9859-9867). We have cloned and expressed the 40-kDa component in Escherichia coli and have purified the recombinant protein to homogeneity. This subunit lacks ATP hydrolytic capacity, but when reconstituted to a 40 kDa-depleted hydrolytic sector, there is a greater than 20-fold increase in calcium-activated, N-ethylmaleimide-sensitive ATP hydrolysis, indicating that this subunit is required for vacuolar-type proton pump function.
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M3 - Article
C2 - 8226880
AN - SCOPUS:0027425070
SN - 0021-9258
VL - 268
SP - 23519
EP - 23523
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 31
ER -