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Lee 1983
Lee 1983
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Printed in Great Britain © 1983 Pergamon Press Ltd
H E X O K I N A S E A N D CARNITINE P A L M I T O Y L T R A N S F E R A S E
ACTIVITIES IN F L I G H T M U S C L E S OF G R A S S H O P P E R
VALANGA NIGRICORNIS(BURM.)
C. W. LEE and C. H. TAN*
Department of Biology, University of Agriculture Malaysia, Serdang,
Selangor, Malaysia
Abstract--1. The activities of hexokinase and carnitine palmitoyltransferase (CPT) activities were
assayed in flight muscle mitochondria of Valanga nioricornis.
2. Kinetic analysis of the enzyme activities yielded the following information: for hexokinase, its Km
(glucose) is 150 #M while its Vm~x is 2.5 #mol/min/mg protein; the corresponding data for CPT are K,,
(palmitoylCoA) of 39/zM and VmaXof 22.2 nmol/min/mg mitochondrial protein.
3. Under conditions of equivalent rates of ATP production the hexokinase-CPT activity ratio is 64,
suggesting that although V. nigricornis is a "mixed" fuel utilizer, it appears that carbohydrate oxidation
constitutes the predominant energy source.
2°l °'10.
J
energy source supporting flight. In other "mixed" fuel
utilizers, variable activity ratios have been reported
(Crabtree & Newsholme, 1975): in the migratory
locusts, L. migratoria and S. gregaria, the ratios are
1.3 and 2.4, respectively; among some species of Lepi-
doptera such as Plusia gamma and Noctua pronuba,
the respective values are 25 and 16. In contrast,
insects that depend exclusively on carbohydrate oxi-
~s. dation have much higher activity ratios: 100 in Perip-
laneta americana (Dictyoptera); 175 in Calliphora
erythrocephala (Diptera); and, 88 in Apis hortorum
0 (Hymenoptera).
0
Pllmitoyl -CoA concentration(uM) The present study has yielded some preliminary
insight into the nature of fuel utilization, hitherto un-
Fig. 1. Carnitine palmitoyltransferase activity in sonicated known, in I/:. nigricornis: the insect may be regarded
flight muscle mitochondria assayed at different concen- as a "mixed" fuel utilizer, but having a predominant
trations of palmitoyl-CoA. Each point represents the mean
capability to oxidise carbohydrate. In the absence of
+ SE for 3 determinations.
information regarding fuel utilization in vivo derived
from flying experiments, this finding is significant.
Moreover, previous attempts to induce flight by the
insect have failed (S. S. Lee and G. J. Goldsworthy,
personal communication).
REFERENCES
BERTHILLER G., COLOBERT L., RICHARD M. & GOT R.
(1970) Glucokinases du foie de rat. Purification et pro-
IIi
prietes des formes particulees. Biochim. biophys. Acta
o
206, 1-16.
CRABTREEI. • NEWSHOLMEE. A. (1972) The activities of
0i lipases and carnitine palmitoyltransferases in muscles
o11 0:2 o~3 d4 o:s from vertebrates and invertebrates. Biochem. J. 130,
o-glucose conclnfrafion (11#t) 697-705.
CRABTREE B. & NEWSHOLME E. A. (1975) Comparative
Fig. 2. Hexokinase activity in crude cytoplasmic extract of aspects of fuel utilization and metabolism by muscle. In
flight muscles: effect of glucose concentration. Each point Insect Muscle (Edited by USHERWOODP. N. R.), pp.
represents mean +SE for 4 determinations. 405-500. Academic Press, London.
DOMROESEK. A. & GILBERTL. I. (1964) The role of lipids
in adult development and flight muscle metabolism in
Goldsworthy, 1976), we have accordingly assayed the Hyalophora cecropia. J. exp. Biol. 41, 573-590.
activity of C P T in flight muscle mitochondria of V. GORNAL A. G., BARDAWILLC. J. & DAVID M. M. (1949)
nigricornis. Figure 1 shows the C P T activities at dif- Determination of serum proteins by means of biuret
reaction. J. biol. Chem. 177, 751.
ferent substrateconcentrations; maximal activity was
JUTSUMA. R. & GOLDSWORTHVG. J. (1976) Fuels for flight
obtainable at 100 #M palmitoyl-CoA. Kinetic analysis in Locusta. J. Insect Physiol. 22, 243-249.
of the data yielded an apparent K s for this substrate KOpEC B. & FRITZ I. B. (1971) Properties of a purified
of 39.2 #m and a Vmax of 22.2 nmol/min/mg mitochon- carnitine palmitoyltransferase, and evidence for the exist-
drial protein. The corresponding data for hexokinase ence of other carnitine acyltransferases. Can. J. Biochem.
are: Km (glucose) of 125 #M, and Vmax of 2.5/~mol/ 49, 941-948.
min/mg protein (Fig. 2). MAYER R. J. & CANDY D. J. (1969) Changes in energy
It is clearly evident that V. nigricornis can poten- reserves during flight of the desert locust, Shistocerca
tially utilise both carbohydrate and lipid as energy gregaria. Comp. Biochem. Physiol. 31,409-418.
SACKTORB. (1975) Utilization of fuels by muscle. In Insect
substrates. A comparison of the two enzyme activities
Biochemistry and Function (Edited by CANDY D. J. &
under equivalent rates of ATP production (Crabtree K[LBY B. A.), pp. 1-81. Chapman and Hall, London.
& Newsholme, 1975), revealed a hexokinase-CPT ac- TAN C. H. & ROBINSONJ. (1982) Activation and transport
tivity ratio of 64. This indicates that although the of fatty acids in ovarian mitochondria: effect of LH.
insect can oxidise lipid, the relatively higher hexo- Endocr. Res. Comm. 9, 1-8.