Jountalof theKoreanChemicalSocieV m, vol.44, No.5
Printedin theRepubficof Korea
Effect of Sperrnine on Phospholipase D Activity in Rat Brain Mitochondrial Preparation
Eun-Hie Koh
Depanment of Chernis~I Duksung Womenk Univers@Seoul132-714, Korea (Received July18, 2000)
~=~ ?5A}3}~~. -&!j]a +x~%}oI]Aj spermin.~1PL~ ~~j~~]~~~ Ca2+,Mg2+2~3 Ban+
spermin#i ?3%+ ++- 7JW71% X= ++93+. * +UJ* % W-7} !iNEQ histamine%-+*
& ~~+lj PL~ =&~ *A] ~+. polyl@’&= Pm ~A#q~] fi~~ ~ 7da]7} ‘~+% ++ =%=
‘}-%-~}%q.7]4
%-4%41*2qAj%
phosphatidylcholine(PCW phosphatidylethanolamine(PE)A}CIM % ~}
CI]%JLoIx]+ tk??q. 61
7]4 ++&+ +% P1=Z=++ PLM PEWAN -’&]a A- +ki+.
ABSTRACT. Phospholipase D (PLD) is the enzyme catalyzing the hydrolysis of the terminaf phosphcester bond of phospholipid head group to produce phosphatidic acid and the conwsponding I&e. The effect of sper- mine on the PLD activity of rat brain mitoehondriaf prepmation was investigated. Spmnine, in the presence of oleic acid activates the rat bmin mitochondrial PLD, whose effect was ftuther enhanced by the presence of divafent cation, Ca2+,Mg2+,or Ba2+.Among the various monoamine testd only histamine at the high con- centration was effective in activation the PLD. Polylysine inc- the PLD activity, particularly, the longer chain of the molecule activated the PLD more effectively. There was no significant difference in the substmite spwificity for the PLD activity between phosphatidylcholine (PC) ,andphoshpatidykthanolamine (PE). This substrate specititiy is different from the PE specificity reported for the intestinal mitochondrial PLD.
–448-
450 R i%M
o 1 2 3 4 5 6
Spermine,mM
Fig. 1. Effect of spermirm on PLD &tivity in brain mitochon- drhl preparation.The PLD nctivity was determined with (0)or without (u) 5 mM oleic acid..
‘~
4ooot
J-m_
Cdrd : I-btamii Lysim mFig. 2. E@cts of vm”ous amines on PLD activity in the mitochond+d preparation. PLD activity was determined with 1 mM amines in the presence of5 rnhf Ca2’.
01 I
o 2 4 6 8 10 12
Hs@rins, mM
Fig. 3.Effect of histamine on PLD activity in themitochon- drial preparation.
/ “-
A0 2U0 400 w 600 1000 1200
Potyiyshe, @l
100O ~
I
o 50 100 150
Polyiysine, @l
Fig. 4. Effects of polylysine on PLD activity in the n3ito- chondrial preparation. A, polyIysine (Av. Mol. VW 1,000> B, polylysine (Av. IMol. WI. 7,000).
Journal of the Korean Chemical Society
024681012 1416 lWdkm, mM
Fig. 5. Effects of divalent metal ions on the spermine-acti- vatedPLD inthe mitoehondml prepamtion. PLD activity was determined at different concentmtions of metal ions in the prmence of 1 mM spermine. CI,Ca2+; $, Mg2+;O, Bal+.
Tile 1. Substrate spedieity of various amineetiects on the PLD activity in mitochondrial preparation
PE Pc PWPC
Activities (cpm) Activities (cpm) Activity Ratio (%)
Spermine lmkl 1761 2232 78.9
Histamine 10 mM 1749 1487 117.6
Polylysine 0.5 mrvl 2M4 2350 87.0
X00. Vol.44 No. 5
452 PFJRM!
i,Psi, J. -K.; Siegel, M. L; Egan, R. W; BiU~ M. .(.
Biol. Chem.. 1988,263, 12472.
2. Exmn, J. H. J. Bzbl, C7zenz 1997,270, 15579.
3.Hmnrmnd, S. M.; ‘Altshuller, Y. M.; Sung, T. -C.;
Rudge, S. A,; Rose, K.; EngebnxhL J.; Morris, A. J.;
Frohmun, M. A. J.BiO1. Chem. 199S, 270,29640.
4. Lee, S. Y.; Yeo, E. -l; Choi, M .-U. Biochem. Bwphy.
Re,s. Comoaut. 1998,244, 825.
5.Meier, K. E.; Gibbs, T. C.; Knoepp, S. M.;Ellz K. M.
BiochenL Biqphy. Acla. 1999. J439, ~99.
6. Mayr, J. A; Kohlwein, S. D.; I%kanf, F. FEBS Lefter 1996,393,236.
7. Madesh, M.;Baksubramanian, K. A. Lipiak 1997,32,
471.
~. L@dus, R. G.; Sokolove.,P. M.J. Bid: Chem. I%, 269, i 8931.
9.Madesh, M.; Brdasubranmian, K. A. )30chem Biophy.
Acts 1997, 1348,324.
10.Jnng, K.; Koh, E.;Choi, M.-V. Ball. Kotwm CheoL SOc. 1989, 10,585.
11. (hi, M. -U.; Abrarnoon,M. B.Biochem Biaphy. ACW 1978,540,337.
12. Kim,B. S.; Hong, S.; Choi, M. -U, Bull. Konm Ciwn.
Sot. 1998, 19/375.
13. Rigobello, M. F?; Bamon, E.; ?&in, O.; Bindnli, A.
Bioclwm Bwphy. Res. Commun. 1995, 217,.144.
14. Made&, M.; Bala..ubramanian,K. A.Biochirn. Biophy.
Acm 1998, 1389,206.
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