Two monoclonal antibody lines directed against subunit IV of cytochrome oxidase: a study of opposite effects.
Two monoclonal lines of antibodies were isolated with specificities against the amino half of Subunit IV of beef heart cytochrome oxidase. The lines had nonoverlapping epitopes. Both bound to the matrix face of membranous oxidase, neither bound to the cytoplasmic face. One line (QA4/C4) stimulated electron transfer in soluble or membranous oxidase, while the other (QA4) inhibited that activity by both oxidase preparations. These effects on electron transfer activity were not altered by the inclusion or omission of detergent. ATP depressed the binding of either antibody to either soluble or membranous oxidase. In the absence of ATP, QA4/C4 stimulated electron transfer only in the high affinity phase of cytochrome c oxidation (with decreased KM and increased Vmax), causing slight inhibition in the low affinity phase (with decreased KM). In the presence of ATP, QA4/C4 abolished the high affinity phase, but did not alter the ATP influence on the low affinity phase. In the absence of ATP, antibodies of line QA4 abolished the low affinity phase, leaving a high affinity phase similar to that induced by ATP. In the presence of ATP, QA4 abolished the high affinity phase, leaving a low affinity phase similar to that seen with ATP alone. This behavior is consistent with the dissection of two catalytic sites for cytochrome c and more than one ATP affector site.
Gai
W Z
WZ
Biology Department, Syracuse University, New York 13244.
Sun
S M
SM
Ding
Y Z
YZ
Freedman
J A
JA
Chan
S H
SH
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000596:Amino Acids / Q000302:isolation & purification; D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D001666:Binding Sites, Antibody; D002417:Cattle; D004579:Electron Transport; D003576:Electron Transport Complex IV / Q000276:immunology / Q000302:isolation & purification*; D000939:Epitopes; D009206:Myocardium / Q000201:enzymology; D010446:Peptide Fragments / Q000302:isolation & purification; D012995:Solubility
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D000939:Epitopes; D010446:Peptide Fragments; D000255:Adenosine Triphosphate; D003576:Electron Transport Complex IV