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Mitochondrial arginine kinase from the heart of the horseshoe crab,Limulus polyphemus

I. Physico-chemical properties and nature of interaction with the mitochondrion

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Summary

Mitochondria from the heart of the horseshoe crab,Limulus polyphemus, contained two distinct activities of arginine kinase (AK). Approximately 50% of AK activity consisted of a fast electrophoretic form (AKmf) which was easily extractable from mitochondria using hypo-tonic buffers. The remaining AK activity was represented by a slow electrophoretic form (AKms) which was tightly bound and could be solubilized only in the presence of non-ionic detergents. Both AKmf and AKms appear to exist in close proximity to the mitochondrial inter-membrane space. AKmf was identical to cytoplasmic AK (AKc) in terms of native electrophoretic mobility, subunit molecular weight, and cyanogen bromide peptide maps. In contrast, AKms had a slightly higher subunit molecular weight and showed some minor, but clear-cut, differences in cyanogen bromide peptide maps from AKmf and AKc. AKms may represent a similar AK form with a hydrophobic targeting sequence for insertion into the mitochondrion and anchoring in the inner membrane. Unlike vertebrate mitochondrial creatine kinase, which binds electrostatically to the mitochondrion, a significant portion of AK inL. polyphemus mitochondria binds via hydrophobic interactions which seems to be characteristic of mitochondrial activities of phosphagen kinases in other invertebrates.

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Abbreviations

AK :

arginine kinase

AK c :

cytoplasmic arginine kinase

AK mf :

fast mitochondrial arginine kinase

AK ms :

slow mitochondrial arginine kinase

ANT :

adenine nucleotide translocase

BSA :

bovine serum albumin

CK :

creatine kinase

CK m :

mitochondrial creatine kinase

CNBr :

cyanogen bromide

DTT :

dithiothreitol

EDTA :

thylene diamine tetraacetic acid

G6PDH :

glucose-6-phosphate dehydrogenase

HAP :

hydroxylapatite

HEPES :

hydroxyethylpiperazine ethanesulfonic acid

HK :

hexokinase

KPi :

potassium phosphate

LDH :

lactate dehydrogenase

MDH :

malate dehydrogenase

MCE :

2-mercaptoethanol

MgAc :

magnesium acetate

MW :

molecular weight

PAGE :

polyacrylamide gel electrophoresis

SDS :

sodium dodecyl sulfate

TRIS :

trishydroxymethylaminomethane

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Authors and Affiliations

  1. Department of Biological Science, Florida State University, 32306, Tallahassee, FL, USA

    C. Doumen & W. R. Ellington

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  1. C. Doumen
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  2. W. R. Ellington
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Doumen, C., Ellington, W.R. Mitochondrial arginine kinase from the heart of the horseshoe crab,Limulus polyphemus . J Comp Physiol B 160, 449–457 (1990). https://doi.org/10.1007/BF01075677

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