Inhibition of Tonoplast ATPase by 2′,3′-Dialdehyde Derivative of ATP

Chow, Wing Ming; Tzeng, Chi Meng; Chen, Chung Shung; Kuo, Soong Yu; Wang, May Yun; Pan, Rong

doi:10.1104/pp.98.1.44

Cited by 4 publications

(4 citation statements)

References 35 publications

(36 reference statements)

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“…5). Therefore, the results with oAMP from this lab were similar to those reported previously by Chow et al (1992) and consistent with the hypothesis that oAMP inhibited the ATPase by acting at or near to the ATP catalytic site. Inhibition of proton transport by oAMP was associated with decreases in the extent of proton transport (data not shown).…”

Section: Effects Of Adpsupporting

confidence: 92%

“…Therefore, inhibition of proton transport by the vacuolar ff-ATPase by ADP was sitnilar to that observed with AMP-PNP and BzATP, but different from the effects of ADP on ATP hydrolysis by the vacuolar H+-ATPase from Beta vulgaris (GrifFith et al 1986). Chow et al (1992) provided convincing evidence that the periodated derivatives of adenosine nucieotides inhibit the tonoplast ATPase by covalently modifying the enzyme at the ATP catalytic site. In the present study the adenosine monophosphate derivative (oAMP) was chosen because results from another study in our laboratory (data not shown) indicated that this compound had better stability and fewer contaminants than the other two periodated derivatives of adenosine nucieotides.…”

Section: Effects Of Adpmentioning

confidence: 97%

“…Nucieotides, AMP-PNP, oAMP and BzATP were obtained from Sigma. Modification of the tonoplast H^-ATPase by oAMP was conducted by the protocol of Chow et al (1992).…”

Section: Preparation Of Inliihitors and Experimental Condltiorsmentioning

confidence: 99%

“…Further evidence for the existence of the catalytic site for ATP on the polypeptide A comes from the research of Chow et al (1992) with tlie 2',3'-dialdehyde derivative of ATP (oATP). The derivative oATP inhibited the tonoplast ATPase from niung bean in time-and concetitrationdependent fashion, consistent with the incorporation of 1 mol of oATP per inactivated ATPase complex.…”

Section: Introductionmentioning

confidence: 99%

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Effects of ATP analogs on the proton pumping by the vacuolar H+-ATPase from maize roots

Bräuer¹,

Tu²

1994

Physiol Plant

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I. 1994. Effects of ATP analogs on the proton pumping by the vacuolar H*-ATPase from maize roots Understanding the regulatory properties of the activities of the 'V-type adenosine triphosphatase (ATPase) on tonoplast memhranes is important in determining the mechanisms by which this enzyme controls cytoplasmic and vacuolar pH. The possible existence of a regulatory site for adeniae nucieotides was examined by comparing the effects of ADP, adenylylimidodiphosphate (AMP-PNP) and 3'-o-(4-benzoy]) benzoyladenine 5'-triphosphate (BzATP) to those of the 2',3'-dialdehyde derivative of AMP (oAMP) and ATP by using highly purified tonoplast vesicles from maize (Zea mays L. cv, FRB 73) roots. The addition of either .^MP-PNP or BzATP reversibly inhibited the initial rate of proton transport catalyzed by the H*-ATPase in a concentration-dependent manner. Le.ss than 20 y.M AMP-PNP or 50 pM BzATP was, sufficient to inhibit half the initial rate of proton transport in the presence of 2 mM ATP and an excess of Mg, Both analogs increased the K,,, for ATP and reduced the maximum enzyme velocity. The presence of ADP also inhibited proton transport. The characteristics of ADP-induced inhibition were similar to those of BzATP and AMP-PNP. The addition of the periodated derivative of AMP (oAMP) irreversibly inhibited the ATPase in a concentration and time-dependent manner similar to that reported previously (Chow et al. 1992, Plant Physiology 98: 44-52). Irreversible inhibition hy oAMP reduced the maximum velocity of the tonoplast ATPase and was prevented by the addition of ATP. The presence of ADP, AMP-PNP or BzATP had no effect on irreversible inhibition by oAMP The effects of ADP, AMP-PNP and BzATP on the kinetics of ATP utilization and the lack of protection against inhibition by oAMP argue in favor of at least two types of nucleotide binding sites on the V-type ATPase from maize root tonoplast membranes.

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Section: Effects Of Adpsupporting

confidence: 92%

Section: Effects Of Adpmentioning

confidence: 97%

“…Nucieotides, AMP-PNP, oAMP and BzATP were obtained from Sigma. Modification of the tonoplast H^-ATPase by oAMP was conducted by the protocol of Chow et al (1992).…”

Section: Preparation Of Inliihitors and Experimental Condltiorsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Effects of ATP analogs on the proton pumping by the vacuolar H+-ATPase from maize roots

Bräuer¹,

Tu²

1994

Physiol Plant

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Inhibition of plant vacuolar H+-ATPase by diethylpyrocarbonate

Chu

Hsiao

Chen

et al. 2001

Biochimica et Biophysica Acta (BBA) - Bioenergetics

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Treatment of the tonoplast H(+)-ATPase from mung bean seedlings (Vigna radiata L.) with histidine-specific modifier, diethyl pyrocarbonate (DEP), caused a marked loss of the ATP hydrolysis activity and the proton translocation in a concentration-dependent manner. The reaction order of inhibition was calculated to be 0.98, suggesting that at least one histidine residue of vacuolar H(+)-ATPase was modified by DEP. The absorbance of the vacuolar H(+)-ATPase at 240 nm was progressively increased after incubation with DEP, suggesting that N-carbethoxyhistidine had been formed. Hydroxylamine, which could break N-carbethoxyhistidine, reversed the absorbance change and partially restored the enzymic activity. The pK(a) of modified residues of vacuolar H(+)-ATPase was kinetically determined to be 6.73, a value close to that of histidine. Thus, it is assuredly concluded that histidine residues of the vacuolar H(+)-ATPase were modified by DEP. Kinetic analysis showed that V(max) but not K(m) of vacuolar H(+)-ATPase was decreased by DEP. This result is interpreted as that the residual activity after DEP inhibition was primarily due to the unmodified enzyme molecules. Moreover, simultaneous presence of DEP and DCCD (N,N'-dicyclohexyl-carbodiimide), an inhibitor modified at proteolipid subunit of vacuolar H(+)-ATPase, did not induce synergistic inhibition, indicating their independent effects. The stoichiometry studies further demonstrate that only one out of four histidine residues modified was involved in the inhibition of vacuolar H(+)-ATPase by DEP. Mg(2+)-ATP, the physiological substrate of vacuolar H(+)-ATPase, but not its analogs, exerted preferentially partial protection against DEP, indicating that the histidine residue involved in the inhibition of enzymatic activity may locate at/or near the active site and directly participate in the binding of the substrate.

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A tryptophan residue involved in the inhibition of plant vacuolar H+-ATPase by 2-hydroxy-5-nitrobenzyl bromide

Kuo¹,

Lin²,

Jiang³

et al. 1998

Functional Plant Biol.

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Treatment of the vacuolar H+ -ATPase from mung bean seedlings (Vigna radiata L.) with the tryptophan modifying agent 2-hydroxyl-5-nitrobenzyl bromide (HNBB), caused a progressive decline of the ATP hydrolysis activity and proton translocation in a time- and concentration-dependent manner. Dithiothreitol could not restore the inhibition of H+ -ATPase by HNBB, indicating possible involvement tryptophan, and not cysteine residues. Protection studies suggested that modified sites might not locate in the active domain. Kinetic analysis shows that Vmax but not Km of H+ -ATPase was changed by HNBB. The reaction order of inactivation by HNBB was calculated as 0.98, implying that at least one tryptophan was labelled. The steady-state dissociation constant (Ki) and the pesudo-first-order rate constant of inhibition (k2) were determined as 1.61 mM and 0.22 min-1, respectively. Furthermore, stoichiometry experiments indicated that 8 mol tryptophan/mol ATPase were modified, when the enzyme activity was completely inhibited. However, a Tsou analysis showed that only one out of these modified tryptophans was crucial to the enzymatic activity. In addition, modification of the vacuolar H+ -ATPase by HNBB led to a decrease in intrinsic fluorescence, suggesting a possible conformational change of the enzyme. Taken together, our data indicate that the tryptophan residue is indispensable to vacuolar H+ -ATPase and the modification of this residue may induce a significant conformational change, consequently resulting in the loss of enzymatic activity.

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Inhibition of Tonoplast ATPase by 2′,3′-Dialdehyde Derivative of ATP

Cited by 4 publications

References 35 publications

Effects of ATP analogs on the proton pumping by the vacuolar H+-ATPase from maize roots

Effects of ATP analogs on the proton pumping by the vacuolar H+-ATPase from maize roots

Inhibition of plant vacuolar H+-ATPase by diethylpyrocarbonate

A tryptophan residue involved in the inhibition of plant vacuolar H+-ATPase by 2-hydroxy-5-nitrobenzyl bromide

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