• BMB Reports
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• 제30권2호
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• pp.157-161
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• 1997

The present paper reports characteristics and specificity of the inhibitory action of $N^{\alpha}-tosyl-L-lysine-chloromethyl\;ketone$ (TLCK) and $N^{\alpha}-tosyl-L-phenylalanine-chloromethyl\;ketone$ (TPCK) on the glucose6-phosphate transporter of rat liver microsomes. The TLCK-induced inhibition was pH dependent. The inhibition constants for TPCK were determined by following pseudo-Lst order reaction mechanism. The inhibition was protected by preincubation with excess amount of glucose-6-phosphate. The results proved that (a) TLCK inactivates the microsomal glucose-6-phosphate transporter, (b) the inhibition results from the modification of sulfhydryl groups of the transporter.

• 한국동물학회지
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• 제33권1호
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• pp.53-62
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• 1990

양서류의 여포난자를 생체외에서 배양하면서 호르몬을 처리하면 난자의 성숙(핵붕괴)을 일으킨다. 본 연구는 북방산개구리의 여포난자를 배양하면서 난자내 단백질 분해효소들의 활성변화를 유도하여 이것이 난자의 성숙에 어떠한 영향을 미치는가를 조사하였다. chymotrypsin의 저해제로 알려진 N$\alpha$ -tosyl-L-phenylalanine-chloromethyl-ketone(TPCK)을 배양액에 처리하면 비교적 낮은 농도(0.001-1 $\mu$M)에서는 호르몬의 도움없이도 난자의 성숙을 유도하나 높은 농도 (100 $\mu$M)에서는 호르몬에 의한 난자의 성숙까지도 억제하는 이중적인 효과를 나타내었다. Trypsin의 저해제인 N$\alpha$ -tosyl-L-phenylalanine-chloromethyl-ketone(TPCK)은 성숙유도능력이 없을 뿐 아니라 progesterone에 의한 난자의 성숙을 억제하였다. Trypsin을 직접 처리했을 때에는 농도에 의존하여(0.001-1$\mu$g/2ml) 호르몬의 도움없이도 난자의 성숙을 유도함을 발견하였다. TLCK나 TPCK의 억제효과는 성숙 초기에만 나타났다. 본 결과는 양서류 난자의 성숙조절 과정에 몇종의 단백질 분해효소들이 참여한다는 것을 시사해주고 있다.

• Fisheries and Aquatic Sciences
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• 제2권2호
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• pp.218-225
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• 1999

A protease without tryptic and chymotryptic activities was purified from the hepatopancreas of shrimp, Penaeus orientalis, using Q-Sepharose ionic exchange, benzamidine Sepharose-6B affinity, Mono-Q, and gel chromatography. Molecular weight (M.W.) of the protease was estimated to be 27kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS­PAGE). The amino acid composition of the protease was different from that of protease from P. japonicus or trypsin from P. orientalis. The protease was completely inhibited by benzamidine, $N\alpha-p-tosyl-L-lysine$ chloromethyl ketone (TLCK), and phenylmethylsulfonyl fluoride (PMSF) and was not affected by leupeptin, pepstatin, N-tosyl-L-phenylalanine chloromethyl ketone (TPCK), iodoacetate, and ethylenediamine tetra acetate (EDTA). The enzyme did not have any activity against Na-benzoyl-DL-arginine p-nitroanilide (BAPNA) or N-benzoyl-L-tyrosine ethyl ester (BTEE) which are specific substrates of trypsin and chymotrypsin, respectively. However, the protease showed hydrolytic activity for a carboxyl terminal of Tyr, Trp, Phe, Glu, and Cys.