Abstract. Introduction. V.I. Lushchak 1,*, T.V. Bagnyukova 1,*, J.M. Storey 2 and K.B. Storey 2

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1 Brzilin Journl of Medicl nd Biologicl Reserch (2001) 34: Effect of exercise on glycolytic enzymes in fish tissues ISSN X 1055 Influence of exercise on the ctivity nd the distriution etween free nd ound forms of glycolytic nd ssocited enzymes in tissues of horse mckerel V.I. Lushchk 1,*, T.V. Bgnyukov 1,*, J.M. Storey 2 nd K.B. Storey 2 1 Deprtment of Nturl Sciences, Precrpthin University, Ivno-Frnkivsk, Ukrine 2 Institute of Biochemistry nd Deprtment of Biology, Crleton University, Ottw, Ontrio, Cnd Correspondence V.I. Lushchk Deprtment of Nturl Sciences Precrpthin University 57 Shevchenko str. Ivno-Frnkivsk, Ukrine E-mil: lushchk@pu.if.u * On leve from Krdg Nturl Reserve, Ntionl Acdemy of Sciences of Ukrine, Kurortne, Feodosi, Crime, Ukrine Received April 17, 2000 Accepted My 15, 2001 Astrct Key words Trchurus mediterrneus Swimming exercise Glycolysis Sucellulr enzyme inding Introduction Brz J Med Biol Res 34(8) 2001

2 1056 V.I. Lushchk et l. Mteril nd Methods Experimentl nimls Biochemicl regents nd coupling enzymes Fish exercise Brz J Med Biol Res 34(8) 2001

3 Effect of exercise on glycolytic enzymes in fish tissues 1057 Preprtion of tissue extrcts Mesurement of enzyme ctivities Brz J Med Biol Res 34(8) 2001

4 1058 V.I. Lushchk et l. Tle 1. Effect of exercise on the ctivities (units/g wet weight) of glycolytic nd ssocited enzymes in white muscle of horse mckerel. Enzyme Control 5-min urst exercise 60-min cruiser exercise HK 0.28 ± ± ± 0.02 PGI 14.5 ± ± ± 0.2 PFK 0.76 ± ± ± 0.04, Ald 52.4 ± ± ± 7.1 PK 241 ± ± ± 7.7 LDH 140 ± ± ± 9 PGM 3.01 ± ± ± 0.32 FBPse 0.57 ± ± ± 0.01, Dt re reported s mens ± SEM of 3-4 determintions in tissue from seprte individuls. P<0.05 compred to the corresponding control vlue nd P<0.05 compred to the 5-min urst exercise group (two-tiled Dunnett test). HK, hexokinse; PGI, phosphoglucoisomerse; PFK, phosphofructokinse; Ald, ldolse; PK, pyruvte kinse; LDH, lctte dehydrogense; PGM, phosphoglucomutse; FBPse, fructose-1,6-isphosphtse. Tle 2. Effect of exercise on the ctivities (units/g wet weight) of glycolytic nd ssocited enzymes in red muscle of horse mckerel. Enzyme Control 5-min urst exercise 60-min cruiser exercise HK 0.42 ± ± ± 0.02 PGI 9.45 ± ± ± 0.36 PFK 0.17 ± ± ± 0.01 Ald 14.2 ± ± ± 1.6 PK 136 ± ± ± 4 LDH 31.7 ± ± ± 1.9 PGM 1.12 ± ± ± 0.05 FBPse 0.55 ± ± ± 0.01 Dt re reported s mens ± SEM of 3-4 determintions in tissue from seprte individuls. P<0.05 compred to the corresponding control vlue nd P<0.05 compred to the 5-min urst exercise group (two-tiled Dunnett test). For revitions, see legend to Tle 1. Results Brz J Med Biol Res 34(8) 2001

5 Effect of exercise on glycolytic enzymes in fish tissues 1059 Tle 3. Effect of exercise on the ctivity (units/g wet weight) of glycolytic nd ssocited enzymes in liver of horse mckerel. Enzyme Control 5-min urst exercise 60-min cruiser exercise HK 0.58 ± ± ± 0.15 PGI 10.8 ± ± ± 0.2 PFK 0.37 ± ± ± 0.09 Ald 2.65 ± ± ± 0.28 PK 6.9 ± ± ± 0.6 LDH 1.57 ± ± ± 0.08 PGM 3.12 ± ± ± 0.28 G6PDH 5.63 ± ± ± 0.04 FBPse 2.88 ± ± ± 0.19 Dt re reported s mens ± SEM of 3-4 determintions in tissue from seprte individuls. P<0.05 compred to the corresponding control vlue nd P<0.05 compred to the 5-min urst exercise group (two-tiled Dunnett test). G6PDH, glucose-6-phosphte dehydrogense. For other revitions, see legend to Tle 1. Tle 4. Effect of exercise on the ctivities (units/g wet weight) of glycolytic nd ssocited enzymes in rin of horse mckerel. Enzyme Control 5-min urst exercise 60-min cruiser exercise HK 0.21 ± ± ± 0.01 PGI 3.72 ± ± ± 0.11 PFK 0.18 ± ± ± 0.01 Ald 1.66 ± ± ± 0.12 PK 36.5 ± ± ± 1.2 LDH 9.20 ± ± ± 0.83 PGM 0.22 ± ± 0.03 G6PDH 0.07 ± ± 0.02 FBPse 0.05 ± Dt re reported s mens ± SEM of 3-4 determintions in tissue from seprte individuls. P<0.05 compred to the corresponding control vlue (two-tiled Dunnett test). G6PDH, glucose-6-phosphte dehydrogense. For other revitions, see legend to Tle 1. Figure 1. Effect of exercise on the distriution of glycolytic nd 60 ssocited enzymes etween 50 free nd ound frctions in white muscle of horse mckerel. Control 40 (white rs), 5-min urst swimming (grey rs) nd 60- min cruiser swimming (lck 30 rs). Dt re reported s 20 mens ± SEM (N = 3-4 individul fishes). P<0.05 compred to the 10 corresponding control vlue nd P<0.05 compred to the 5-min 0 HK PGI PFK Ald PK LDH PGM FBP urst exercise group (two-tiled Dunnett test). HK, hexokinse; PGI, phosphoglucoisomerse; PFK, phosphofructokinse; Ald, ldolse; PK, pyruvte kinse; LDH, lctte dehydrogense; PGM, phosphoglucomutse; FBP, fructose-1,6-isphosphtse. Bound enzyme (%) Brz J Med Biol Res 34(8) 2001

6 1060 V.I. Lushchk et l. Figure 2. Effect of exercise on the distriution of enzymes etween free nd ound frctions in red muscle of horse mckerel. Other informtion s in the legend to Figure 1. Figure 3. Effect of exercise on the distriution of enzymes etween free nd ound frctions in liver of horse mckerel. G6PDH, glucose-6-phosphte dehydrogense. Other informtion s in the legend to Figure 1. Figure 4. Effect of exercise on the distriution of enzymes etween free nd ound frctions in rin of horse mckerel. Other informtion s in the legend to Figure 1. Bound enzyme (%) Bound enzyme (%) Bound enzyme (%) HK PGI PFK Ald HK PGI PFK Ald PK LDH PGM FBP PK LDH PGM FBP G6PDH,,, HK PGI PFK Ald PK LDH Discussion Brz J Med Biol Res 34(8) 2001

7 Effect of exercise on glycolytic enzymes in fish tissues 1061 Brz J Med Biol Res 34(8) 2001

8 1062 Brz J Med Biol Res 34(8) 2001 V.I. Lushchk et l.

9 Effect of exercise on glycolytic enzymes in fish tissues 1063 Acknowledgments References 1. Shulmn GE (Editor) (1978). Elements of Physiology nd Biochemistry of Generl nd Active Metolism in Fishes. Nukov Dumk, Kyiv. 2. Rhim ZHA, Luty G & Griffiths JR (1979). Activtion of AMP minohydrolse during skeletl-muscle contrction. Biochemicl Journl, 184: Wokom A & Johnston IA (1983). Aneroic metolism during ctivity in rinow trout (Slmo girdneri ). Experienti, 39: Prkhouse WS, Doson GP, Belcstro AN & Hochchk PW (1987). The role of intermediry metolism in the mintennce of proton nd chrge lnce during exercise. Moleculr nd Cellulr Biochemistry, 77: Prkhouse WS, Doson GP & Hochchk PW (1988). Control of glycogenolysis in rinow trout muscle during exercise. Cndin Journl of Zoology, 66: Mommsen TP & Hochchk PW (1988). The purine nucleotide cycle s two temporlly seprted metolic units: study on trout muscle. Metolism, 37: Vn den Thillrt G, Vn Wrde A, Muller HJ, Erkelens C, Addink A & Lugtenurg J (1989). Fish muscle energy metolism mesured y in vivo 31 P-NMR during noxi nd recovery. Americn Journl of Physiology, 256: R922-R Storey KB (1991). Metolic consequences of exercise in orgns of rinow trout. Journl of Experimentl Zoology, 260: Schulte PM, Moyes CD & Hochchk PW (1992). Integrting metolic pthwys in post-exercise recovery of white muscle. Journl of Experimentl Biology, 166: Lushchk VI (1995). Role of phosphoryltion nd redistriution of glycolytic enzymes in dpttion of hydroiontes to enviromentl conditions. Hydroiologicl Journl, 31: Msters CJ, Reid S & Don M (1987). Glycolysis - new concepts in n old pthwy. Moleculr nd Cellulr Biochemistry, 76: Brooks SPJ & Storey KB (1988). Sucellulr enzyme inding in glycolytic control: in vivo studies with fish muscle. Americn Journl of Physiology, 255: R289-R Lushchk VI (1996). AMP-deminse: functions, moleculr properties nd locliztion in the cell. Biochemistry (Moscow), 61: Lushchk VI (1996). Prticiption of suprmoleculr complex formtion in the regultion of glycolytic enzymes. Ukrinin Biochemicl Journl, 68: Hsu SC & Moldy RS (1990). Glycerldehyde-3-phosphte dehydrogense is mjor protein ssocited with the plsm memrne of retinl photoreceptor outer segments. Journl of Biologicl Chemistry, 265: Lushchk VI (1991). Properties of memrne-ound lctte dehydrogense from white skte muscle. Biokhimii, 56: Brooks SPJ & Storey KB (1993). Control of glycolytic enzyme inding: effect of chnging enzyme sustrte concentrtions on in vivo enzyme distriution. Moleculr nd Cellulr Biochemistry, 122: Luther MA & Lee JC (1986). The role of phosphoryltion in the interction of rit muscle phosphofructokinse with F- ctin. Journl of Biologicl Chemistry, 261: Kurgnov BI (1986). The role of multienzyme complexes in integrtion of cellulr metolism. Journl of Theoreticl Biology, 119: Ehmnn JD & Hultin HO (1973). Sustrte inhiition of solule nd ound lctte dehydrogense (isoenzyme 5). Archives of Biochemistry nd Biophysics, 154: Lushchk VI (1992). Lctte dehydrogense interction with the structurl cell components: the possile physiologicl significnce. Biokhimii, 57: Lushchk VI (1992). Free nd memrneound lctte dehydrogense from white driving muscles of skte. Biochemistry Interntionl, 26: Liou R-S & Anderson S (1980). Activtion of rit muscle phosphofructokinse y F-ctin nd reconstituted thin filments. Biochemistry, 19: Kuo H-J, Mlencik DA, Liou RS & Anderson SR (1986). Fctors ffecting the ctivtion of rit muscle phosphofructokinse y ctin. Biochemistry, 25: Lushchk VI (1993). Free nd ound pyruvte kinse from fish rin: properties nd redistriution fter hypoxi. Biochemistry nd Moleculr Biology Interntionl, 29: Brz J Med Biol Res 34(8) 2001

10 1064 V.I. Lushchk et l Clrke FM, Shw FD & Morton DJ (1980). Effect of electricl stimultion post mortem of ovine muscle on the inding of glycolytic enzymes. Functionl nd structurl implictions. Biochemicl Journl, 186: Brooks SPJ & Storey KB (1991). Studies on the regultion of enzyme inding during noxi in isolted tissues of Busycon cnlicultum. Journl of Experimentl Biology, 156: Duncn JA & Storey KB (1991). Role of enzyme inding in muscle metolism of the goldfish. Cndin Journl of Zoology, 69: Lushchk VI & Storey KB (1994). Effect of exercise on the distriution of enzymes in trout white muscle nd kinetic properties of AMP-deminse from free nd ound frctions. Fish Physiology nd Biochemistry, 13: Lushchk VI, Bhnjukov TV & Storey KB (1998). Effect of hypoxi on the ctivity nd inding of glycolytic nd ssocited enzymes in se scorpion tissues. Brzilin Journl of Medicl nd Biologicl Reserch, 31: Booth FW & Thompson DB (1991). Moleculr nd cellulr dpttion of muscle in response to exercise: perspectives of vrious models. Physiologicl Reviews, 71: Storey KB & Brooks SPJ (1995). Is glycolytic rte controlled y the reversile inding of enzymes to sucellulr structures? In: Hochchk PW & Mommsen TP (Editors), Biochemistry nd Moleculr Biology of Fishes. Vol. 4. Elsevier, Amsterdm, Depinto V, Aljml JA & Plmieri F (1993). Hexokinse-inding protein. Loction of the dicyclohexylcrodiimide-rective glutmte residue in the ovine hert mitochondril porin. Journl of Biologicl Chemistry, 268: Hssinen IE & Vuokil PT (1993). Hexokinse-inding protein. Rection of dicyclohexylcrodiimide with mitochondril proteins. Biochimic et Biophysic Act, 1144: Brz J Med Biol Res 34(8) 2001