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Physical properties and metabolite regulation of ribulose bisphosphate carboxylase from Thiobacillus A2

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Abstract

Purified ribulose-bisphosphate carboxylase (EC 4.1.1.39) was strongly and equally inhibited either by ADP or GDP and to a lesser extent by IDP. AMP or ATP exerted little effect on activity. Inhibition by the nucleotide diphosphates was competitive with respect to RuBP and non-competitive with respect to “CO2” and Mg2+, respectively. Treatment of the enzyme with urea or guanidine-HCl resulted in rapid loss of activity that was not restored by dialysis even in the presence of Mg2+ and cysteine. Sodium dodecyl sulfate electrophoresis of 8.0 M urea treated enzyme revealed the presence of a fast-moving (small) sub-unit with molecular weight 14150 and a slower moving (large) sub-unit with molecular weight 68000. Examination of native enzyme by sodium dodecyl sulfate electrophoresis gave sub-units of 13700 and 55500 respectively. The amino acid content standardized to phenylalanine was essentially similar to that from other sources. Arrhenius plots showed a “break” at 29°C with an E a of 12.34 kcal per mole for the steeper part of the curve and a ΔH of 11.43 kcal per mole while for the less steep region, the E a was 1.04 kcal per mole and the ΔH 1.92 kcal per mole.

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Abbreviations

ADP:

adenosine-5′-diphosphate

AMP:

adenosine-5′-monophosphate

ATP:

adenosine-5′-triphosphate

CDP:

cytidine-5′-diphosphate

CMP:

cytidine-5′-monophosphate

CTP:

cytidine-5′-triphosphate

FDP:

fructose-1,6′-diphosphate

F6P:

fructose-6-phosphate

GDP:

guanosine-5′-diphosphate

GMP:

guanosine-5′-monophosphate

G6P:

glucose-6-phosphate

GTP:

guanosine-5′-triphosphate

IDP:

inosine-5′-diphosphate

IMP:

inosine-5′-monophosphate

PEP:

phosphoenolpyruvate

6PG:

6-phosphogluconate

R1P:

ribose-1-phosphate

R5P:

ribose-5-phosphate

RuBP:

ribulose-1,5-bisphosphate

SDS:

sodium dodecyl sulfate

TDP:

thymidine-5′-diphosphate

TMP:

thymidine-5′-monophosphate

TTP:

thymidine-5′-triphosphate

UDP:

uridine-5′-diphosphate

UMP:

uridine-5′-monophosphate

UTP:

uridine-5′-triphosphate

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Authors and Affiliations

  1. Department of Biology, University of Waterloo, N2L 3G1, Waterloo, Ontario, Canada

    A. M. Charles & B. White

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  1. A. M. Charles
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  2. B. White
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Charles, A.M., White, B. Physical properties and metabolite regulation of ribulose bisphosphate carboxylase from Thiobacillus A2 . Arch. Microbiol. 108, 203–209 (1976). https://doi.org/10.1007/BF00428952

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