Relationship of Tightly Bound ADP and ATP to Control and Catalysis by Chloroplast ATP Synthase

Jun mei Zhou, Zhixiong Xue, Ziyun Du, Teri Melese, Paul D. Boyer

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Abstract

Whether the tightly bound ADP that can cause a pronounced inhibition of ATP hydrolysis by the chloroplast ATP synthase and F 1 ATPase (CF 1 )is bound at catalytic sites or at noncatalytic regulatory sites or both has been uncertain. We have used photolabeling by 2-azido-ATP and 2-azido-ADP to ascertain the location, with Mg 2+ activation, of tightly bound ADP (a) that inhibits the hydrolysis of ATP by chloroplast ATP synthase, (b) that can result in an inhibited form of CF 1 that slowly regains activity during ATP hydrolysis, and (c) that arises when low concentrations of ADP markedly inhibit the hydrolysis of GTP by CF l. The data show that in all instances the inhibition is associated with ADP binding without inorganic phosphate (P i ) at catalytic sites. After photophosphorylation of ADP or 2-azido-ADP with [ 32 P]P j , similar amounts of the corresponding triphosphates are present on washed thylakoid membranes. Trials with appropriately labeled substrates show that a small portion of the tightly bound 2-azido-ATP gives rise to covalent labeling with an ATP moiety at noncatalytic sites but that most of the bound 2-azido-ATP gives rise to covalent labeling by an ADP moiety at a catalytic site. We also report the occurrence of a 1-2-min delay in the onset of the Mg 2+ -induced inhibition after addition of CF 1 to solutions containing Mg 2+ and ATP, and that this delay is not associated with the filling of noncatalytic sites. A rapid burst of P i formation is followed by a much lower, constant steady-state rate. the burst is not observed with GTP as a substrate or with Ca 2+ as the activating cation.

Original languageEnglish (US)
Pages (from-to)5129-5135
Number of pages7
JournalBiochemistry
Volume27
Issue number14
DOIs
StatePublished - Jul 1 1988

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Chloroplast Proton-Translocating ATPases
Catalysis
Adenosine Diphosphate
Adenosine Triphosphate
Hydrolysis
Catalytic Domain
Guanosine Triphosphate
Labeling
Photophosphorylation
Regain
Thylakoids
Proton-Translocating ATPases
Substrates
Cations
Chemical activation
Phosphates
Membranes

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Relationship of Tightly Bound ADP and ATP to Control and Catalysis by Chloroplast ATP Synthase. / Zhou, Jun mei; Xue, Zhixiong; Du, Ziyun; Melese, Teri; Boyer, Paul D.

In: Biochemistry, Vol. 27, No. 14, 01.07.1988, p. 5129-5135.

Research output: Contribution to journalArticle

Zhou, Jun mei ; Xue, Zhixiong ; Du, Ziyun ; Melese, Teri ; Boyer, Paul D. / Relationship of Tightly Bound ADP and ATP to Control and Catalysis by Chloroplast ATP Synthase. In: Biochemistry. 1988 ; Vol. 27, No. 14. pp. 5129-5135.
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