TY - JOUR
T1 - Extraction, Purification, and Reconstruction of the Chloroplast N,N′-Dicyclohexylcarbodiimide-Binding Proteolipid
AU - Azzi, Angelo
AU - Sigrist-Nelson, Kristine
AU - Nelson, Nathan
N1 - Funding Information:
This work was supported by a grant from the United States-Israel Binational Science Foundation, Jerusalem, Israel.
PY - 1986/1/1
Y1 - 1986/1/1
N2 - This chapter describes the techniques for the extraction, purification, and reconstruction of the chloroplast N, N'-dicyclohexylcarbodiimide-binding proteolipid (DCCD). The techniques described are suitable for isolating and reconstituting in an active form the proteolipid subunit of chloroplast ATPase. The isolation is initiated by a one-step single-phase l-butanol extraction. The ATPase complex of chloroplasts, mitochondria, and bacterial membranes is composed of two distinct parts: F1, and F0. The identification of F0 is done by the use of DCCD, a reagent that inhibits ATPase activity of the F0-F1 complex, but not that of the F1 alone. DCCD also reduces the proton permeability of the membrane from which F1 is removed, suggesting that the action of the inhibitor is to block a site(s) of the F0 that participates in proton translocation. DCCD binds only to one of the F0 subunits, called as DCCD-binding proteolipid, or the proteolipid.
AB - This chapter describes the techniques for the extraction, purification, and reconstruction of the chloroplast N, N'-dicyclohexylcarbodiimide-binding proteolipid (DCCD). The techniques described are suitable for isolating and reconstituting in an active form the proteolipid subunit of chloroplast ATPase. The isolation is initiated by a one-step single-phase l-butanol extraction. The ATPase complex of chloroplasts, mitochondria, and bacterial membranes is composed of two distinct parts: F1, and F0. The identification of F0 is done by the use of DCCD, a reagent that inhibits ATPase activity of the F0-F1 complex, but not that of the F1 alone. DCCD also reduces the proton permeability of the membrane from which F1 is removed, suggesting that the action of the inhibitor is to block a site(s) of the F0 that participates in proton translocation. DCCD binds only to one of the F0 subunits, called as DCCD-binding proteolipid, or the proteolipid.
UR - http://www.scopus.com/inward/record.url?scp=77957067381&partnerID=8YFLogxK
U2 - 10.1016/S0076-6879(86)26054-1
DO - 10.1016/S0076-6879(86)26054-1
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AN - SCOPUS:77957067381
SN - 0076-6879
VL - 126
SP - 520
EP - 527
JO - Methods in Enzymology
JF - Methods in Enzymology
IS - C
ER -