Cryoenzymologic Studies on Arginine Kinase: Solvent, Temperature and pH Effects on the Overall Reaction

Travers, Franck; Bertrand, Raoul; Roseau, Gilbert; Thoại, Nguyễn Văn

doi:10.1111/j.1432-1033.1978.tb12478.x

Cited by 23 publications

(21 citation statements)

References 18 publications

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“…The complexity of the system is further increased by lowering the temperature as under these conditions there is a change in a ratelimiting step (see below). Although the above results are less precise and the interpretation less clear, there could be a parallel between these and our previous results with arginine kinase [7]. Thus, because of the observed effect of the solvent, the first p K could be anionic and correspond to the pK of the phosphate group transferred.…”

Section: The Effects Of Ethylene Glycol On the Myosin S L Ph Profilesmentioning

confidence: 45%

“…The reason for the observed breaks is, therefore, simple: it is caused by a change in the rate-limiting step. This explanation is a consequence of thermodynamic reasoning and it has already been exploited in the case of arginine kinase [7]. We also studied the acto-Mg ATPase activity of myosin.…”

Section: Temperature Dependence Of Kmentioning

confidence: 99%

“…4). Although this phenomenon also occurs with other systems (for example arginine kinase [7]) and reasonable explanations have been proposed in certain cases [20], there is as yet no general interpretation. This progressive and reversible inhibition by an organic solvent remains, therefore, an experimental fact which could be caused by an effect of the solvent on the active site of myosin as well as on the substrate itself.…”

Section: Ejfect Of Ethylene Glycol On the Steady-state Activity Of Mymentioning

confidence: 99%

“…As already indicated in a preceding publication on arginine kinase [7], cryoenzymological studies should be preceded by a detailed investigation of the influence of the unusual conditions used (organic solvent and low temperatures) on the overall reaction under study. This is the object of the present work.…”

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confidence: 99%

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Cryoenzymological Studies on Myosin Subfragment 1

Travers¹,

Hillaire²

1979

European Journal of Biochemistry

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The ATPase activities of myosin subfragment 1 were studied under subzero conditions with ethylene glycol as the antifreeze.The pH profile of the Ca ATPase activity was affected by both ethylene glycol and the temperature but the pH profile of the Mg ATPase activity was only slightly affected.At 20 "C the Mg ATPase activity was not affected by the solvent. The Ca and acto-Mg ATPase activities decreased to similar extents as the solvent concentration was increased. These results are discussed in terms of the accessibility of the active site.The Mg, Ca and acto-Mg ATPase activities were studied down to -20 "C, in 40 % ethylene glycol. All three systems showed an increase in activation energy at low temperatures. These results were compared to previous work and they were analysed in terms of a change in the rate-limiting step.When myosin S1 is mixed with ATP, difference spectra are obtained whose amplitudes decrease with time and which are a manifestation of the intermediate M**. ADP . Pi (the product complex).

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Section: The Effects Of Ethylene Glycol On the Myosin S L Ph Profilesmentioning

confidence: 45%

Section: Temperature Dependence Of Kmentioning

confidence: 99%

Section: Ejfect Of Ethylene Glycol On the Steady-state Activity Of Mymentioning

confidence: 99%

mentioning

confidence: 99%

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Cryoenzymological Studies on Myosin Subfragment 1

Travers¹,

Hillaire²

1979

European Journal of Biochemistry

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“…The answer is probably not. First, as found for arginine kinase [16], the K, values for ATP and guanidine substrate were not significantly affected by these conditions [ I ] . Second, the energies of activation for the overall catalytic reactions were almost identical in pure water and in 40% ethylene glycol [ I , 161.…”

Section: Discussionmentioning

confidence: 99%

Cryoenzymic Studies on the Transition‐State Analog Complex Creatine Kinase ‐ ADPMg ‐ Nitrate ‐ Creatine

Travers

Barman

1980

European Journal of Biochemistry

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Concomitant with the formation of the transition state analog complex creatine kinase . ADPMg . nitrate . creatine [Milner-White, E. J. and Watts, D. C. (1971) Bioclzem. J. 122, 727-7401 there results a large difference spectrum. The shape of the spectrum was characteristic of tryptophan exposure.The kinetics of formation and final amplitude of the spectrum were studied at -15 "C using a stopped-flow apparatus. The results obtained allowed for a plausible reaction pathway for nitrate fixation: the ordered addition of nitrate and creatine to the binary enzyme-ADPMg complex, a slow protein isomerization and the final addition of a molecule each of nitrate and creatine.The kinetic parameters for the formation of the transition state analog complex were compared with those already known for the overall reaction obtained under the same conditions [Barman, T. E., Brun, A. and Travers, F. (1980) Eur. J. Biochem. 110, 397-4031, This comparison revealed a striking analogy between the two processes. This suggests that the conformation of creatine kinase in the analog complex is very similar to that in the transition state complex on the catalytic pathway.In the preceding paper [I] we used the rapid flowquench apparatus under subzero conditions to search for intermediates in the creatine kinase reaction pathway. Under these conditions the enzyme appeared to be partitioned exclusively between enzyme-substrate and enzyme-product complexes : we could not detect any phosphoenzyme or labile phosphate intermediate down to 4ms. We obtained a complex product (creatine phosphate) progress curve which allowed us to estimate the rate constants pertaining to a simple mechanism. These results make it likely that creatine kinase proceeds via a direct in-line transfer between the participating substrates, as has already been suggested by Milner-White and Watts [2].Another approach towards the study of enzyme mechanisms is to use transition state analogs [3,4]. In the case of creatine kinase there is no simple, single analog inhibitor. However, Milner-White and Watts [2] showed that the combination MgADP, nitrate and creatine is a powerful inhibitor of the enzyme. They suggest that the nitrate binds to the enzyme at the site normally occupied by the transAbbreviations. Cr, creatine; CrP, creatine phosphate. Enzymes. Creatine kinase (EC 2.7.3.2); arginine kinase (EC 2.7.3.3). ferable phosphate and they conclude that the complex enzyme . ADPMg . NO3 . creatine may mimic the putative transition state complex enzyme . ADPMg . PO4 .creatine. Now, such a complex would occupy a position between an enzyme-substrate and an enzymeproduct complex on the overall catalytic reaction. Since we already have information on the rate constants involved in this step, it seemed to us important to investigate further the formation of the enzyme . ADPMg . NO3 . Cr (nitrate complex) and to see what analogies there might be between the two processes.In this paper we study the formation of the creatine kinase . ADPMg ' NOs ' Cr complex at subzero temperatures [...

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Cold-adapted Features of Arginine Kinase from the Deep-sea Clam Calyptogena kaikoi

et al. 2011

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The heterodont clam Calyptogena kaikoi, which inhabits depths exceeding 3,500 m where low ambient temperatures prevail, has an unusual two-domain arginine kinase (AK) with molecular mass of 80 kDa, twice that of typical AKs. The purpose of this work is to investigate the nature of the adaptations of this AK for functioning at low temperatures. Recombinant C. kaikoi AK constructs were expressed, and their two-substrate kinetic constants (k(cat), K(a), and K(ia)) were determined at 10°C and 25°C, respectively. When measured at 25°C, the K(ia) values were tenfold larger than those for corresponding K(a) values, while at 10°C, the K(ia) values decreased remarkably, but the K (a) values were almost unchanged. The Calyptogena two-domain enzyme has threefold higher catalytic efficiency, calculated by k (cat)/(K(a)(ARG)·K(ia)(ATP) ), at 10°C, than that at 25°C, reflecting adaptation for function at reduced ambient temperatures. The activation energy (E(a)) and thermodynamic parameters were determined for Calyptogena two-domain enzyme and compared with those of two-domain enzymes from mesophilic Corbicula and Anthopleura. The value for E(a) of Calyptogena enzyme were about half of those for mesophilic enzymes, and a larger decrease in entropy was observed in Calyptogena AK reaction. Although large decrease in entropy increases the ΔG(o‡) value and consequently lowers the k(cat) value, this is compensated with its lower E(a) value thereby minimizing the reduction in its k(cat) value. These thermodynamic properties, together with the kinetic ones, are also present in the separated domain 2 of the Calyptogena two-domain enzyme.

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Cryoenzymologic Studies on Arginine Kinase: Solvent, Temperature and pH Effects on the Overall Reaction

Cited by 23 publications

References 18 publications

Cryoenzymological Studies on Myosin Subfragment 1

Cryoenzymological Studies on Myosin Subfragment 1

Cryoenzymic Studies on the Transition‐State Analog Complex Creatine Kinase ‐ ADPMg ‐ Nitrate ‐ Creatine

Cold-adapted Features of Arginine Kinase from the Deep-sea Clam Calyptogena kaikoi

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