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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

Journal of Comparative Physiology B volume 160, pages 449–457 (1990)Cite this article

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

References

  • Bessman SP, Carpenter CL (1985) The creatine-creatine phosphate energy shuttle. Ann Rev Biochem 65:831–862

    Google Scholar 

  • Blethen SL, Kaplan NO (1968) Characteristics of arthropod arginine kinases. Biochem 7:2123–2135

    Google Scholar 

  • Brooks SPJ, Suelter CH (1987) Association of chicken mitochondrial creatine kinase with the inner mitochondrial membrane. Arch Biochem Biophys 253:122–132

    PubMed  Google Scholar 

  • Chen C-H, Lehninger AL (1973) Respiration and phosphorylation by mitochondria from the hepatopancreas of the blue crab (Callinectes sapidus). Arch Biochem Biophys 154:449–459

    PubMed  Google Scholar 

  • Cheneval D, Carafoli E (1988) Identification and primary structure of the cardiolipin-binding domain of mitochondrial creatine kinase. Eur J Biochem 172:1–9

    PubMed  Google Scholar 

  • Chih CP, Ellington WR (1987) Studies on the mitochondria and enzymes from the ventricle of the whelk,Busycon contrarium. Comp Biochem Physiol 86B(3):541–545

    Google Scholar 

  • Doonan S, Barra D, Bossa F (1984) Structural and genetic relationships between cytosolic and mitochondrial isoenzymes. Int J Biochem 16:1193–1199

    PubMed  Google Scholar 

  • Doumen C, Ellington WR (1987) Arginine phosphokinase activity in mitochondria from the tubular heart of the horseshoe crab,Limulus polyphemus. Am Zool 27(4):54A

    Google Scholar 

  • Doumen C, Ellington WR (1989) Substrate preferences of the heart mitochondria of the horseshoe crab,Limulus polyphemus. Comp Biochem Physiol 93B:883–887

    Google Scholar 

  • Doumen C, Ellington WR (1990) Mitochondrial arginine kinase from the horseshoe crabLimulus polyphemus. II. Catalytic properties and studies of potential functional coupling with oxidative phosphorylation. J Comp Physiol B 160

  • Hird FJR, Robin A (1985) Studies on phosphagen synthesis by mitochondrial preparations. Comp Biochem Physiol 80B(3):517–520

    Google Scholar 

  • Hochachka PW, Fields JHA, Mommsen TP (1983) Metabolic and enzyme regulation during rest-to-work transition: a mammal vs mollusc comparison. In Hochachka PW (ed) The mollusca Vol. 1. Academic Press, New York, pp 56–89

    Google Scholar 

  • Ieyengar MR, Ieyengar CL (1980) Interaction of creatine kinase isoenzymes with beef heart mitochondrial membrane: a model for association of mitochondrial and cytoplasmic isoenzymes with inner membrane. Biochem 19:2176–2182

    Google Scholar 

  • Jacobs H, Heldt HW, Klingenberger M (1964) High activity of creatine kinase in mitochondria from muscle and brain and evidence for a separate mitochondrial isoenzyme of creatine kinase. Biochem Biophys Res Comm 16(6):516–521

    PubMed  Google Scholar 

  • Jacobus WE (1985) Respiratory control and the integration of high-energy phosphate metabolism by mitcohondrial creatine kinase. Ann Rev Physiol 47:707–725

    Google Scholar 

  • Jacobus WE, Lehninger AL (1973) Creatine kinase of rat heart mitochondria: coupling of creatine kinase phosphorylation to electron transport. J Biol Chem 248:4803–4810

    PubMed  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of Bacteriophage T4. Nature (London) 227:680–685

    Google Scholar 

  • Lowry OH, Rosenbrough MJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    PubMed  Google Scholar 

  • Morrisson JF (1973) Arginine kinase and other invertebrate guanidino kinases. In: Boyer PD (ed) The enzymes. Academic Press, New York, pp 457–486

    Google Scholar 

  • Meyer MA, Sweeney HL, Kushmerick MJ (1984) A simple analysis of the “phosphocreatine shuttle”. Am J Physiol 246:C365–377

    PubMed  Google Scholar 

  • Muller M, Moser R, Cheneval D, Carafoli E (1985) Cardiolipin is the membrane receptor for mitochondrial creatine phosphokinase. J Biol Chem 260(6):3839–3843

    PubMed  Google Scholar 

  • Munneke LR, Collier GE (1985) Mitochondrial arginine kinase inDrosophila melanogaster. Genetics 110:s85

  • Munneke LR, Collier GE (1988) Cytoplasmic and mitochondrial arginine kinases inDrosophila: evidence for a single gene. Biochem Gen 26:131–141

    Google Scholar 

  • Newsholme EA, Beis I, Leech AR, Zammit VA (1978) The role of creatine kinase and arginine kinase in muscle. Biochem J 172:533–537

    PubMed  Google Scholar 

  • Nikodem V, Fresco JR (1979) Protein fingerprinting by SDS-gel electrophoresis after partial fragmentation with CNBr. Anal Biochem 97:382–386

    PubMed  Google Scholar 

  • O'Hare MC, Doonan S (1985) Purification and structural comparisons of the cytosolic and mitochondrial isoenzymes of fumarase from pig liver. Biochim Biophys Acta 82:127–134

    Google Scholar 

  • Saks VA, Chernousova GB, Voronkov Iul, Smirnov VN, Chazov EI (1974) Study of energy transport mechanism in myocardial cells. Circ Res 34–35 (sup II.III):138–148

    Google Scholar 

  • Saks VA, Kupriyanov VV, Elizarova GV, Jacobus WE (1980) Studies of energy transport in heart cells. The importance of creatine kinase localization for the coupling of mitochondria phosphorylcreatine production to oxidative phosphorylation. J Biol Chem 255(2):755–763

    PubMed  Google Scholar 

  • Saks VA, Kuznetsov AV, Kupriyanov VV, Mecili MV, Jacobus WE (1985) Creatine kinase of rat heart mitochondria. The demonstration of functional coupling to oxidative phosphorylation in an inner membrane-matrix preparation. J Biol Chem 260(12):7757–7764

    PubMed  Google Scholar 

  • Schatz G (1987) Signals guiding proteins to their correct locations in mitochondria. Eur J Biochem 165:1–6

    PubMed  Google Scholar 

  • Schlegel J, Zurbriggen B, Wegmann G, Wyss M, Eppenberger HM, Walliam T (1988) Native mitochondrial creatine kinase forms octameric structures. I: Isolation of two interconvertible mitochondrial creatine kinase forms, dimeric and octameric mitochondrial creatine kinase: characterization, localization and structure-function relationships. J Biol Chem 263:16942–16953

    PubMed  Google Scholar 

  • Schnyder T, Engel A, Lustig A, Walliman T (1988) Native mitochondrial creatine kinase forms octameric structures. II: Characterization of dimers and octamers by ultracentrifugation, direct mass measurements by scanning transmission electron microscopy, and image analysis of single mitochondrial creatine kinase octamers. J Biol Chem 263:16954–16962

    PubMed  Google Scholar 

  • Sharov VG, Saks VA, Smirnov VN, Chazov EI (1977) An electron microscopic histochemical investigation of the localization of creatine phosphokinase in heart cells. Biochim Biophys Acta 468:495–501

    PubMed  Google Scholar 

  • Skorkowski EF, Aleksandrowicz Z, Wrzolkowa T, Swierczynski J (1976) Isolation and some properties of mitochondria from the abdomen muscle of the crayfish,Orconectes limosus. Comp Biochem Physiol 55B:493–500

    Google Scholar 

  • Storey KB (1977) Purification and characterization of arginine kinase from the mantle muscle of the squid,Symplectoteuthis oualaniensis. Arch Biochem Biophys 179:518–526

    PubMed  Google Scholar 

  • Tombes RM, Shapiro BM (1985) Metabolic channeling: a phosphorylcreatine shuttle to mediate high energy phosphate transport between sperm mitochondrion and tail. Cell 41:325–334

    PubMed  Google Scholar 

  • Tombes RM, Shapiro BM (1987) Enzyme termini of a phosphocreatine shuttle. Purification and characterization of two creatine kinase isozymes from sea urchin sperm. J Biol Chem 262(33):16011–16019

    PubMed  Google Scholar 

  • Turner DC, Wallimann T, Eppenberger HM (1973) A protein that binds specifically to the M-line of skeletal muscle is identified as the muscle form of creatine kinase. Proc Natl Acad Sci USA 70:702–707

    PubMed  Google Scholar 

  • Vial C, Font B, Goldschmidt D, Gauteron DC (1979) Dissociation and reassociation of creatine kinase with heart mitochondria; pH and phosphate dependence. Biochim Biophys Acta 88:1352–1359

    Google Scholar 

  • Wickner WT, Lodish HF (1985) Multiple mechanisms of protein insertion into and across membranes. Science 230:400–407

    PubMed  Google Scholar 

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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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  • DOI: https://doi.org/10.1007/BF01075677

Key words

  • Arginine kinase
  • Horseshoe crab
  • Mitochondria
  • Compartmentation
  • ATP-buffering