Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Effect of Replacing Glutamate-219 with Glutamine or Alanine in M u ri ne Mono-AD P-ri bosyltra nsferase

Murine mono-ADP-ribosyltransferase에서 glutamic acid-219를 glutamine혹은 alanine 으로의 치환에 의한 효과

  • 김현주 (울산대학교 생명과학부)
  • Published : 1998.04.01
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Abstract

Two distinct ADP-ribosyltransferases, termed Yac-1 and Yac-2 from mouse lymphoma cells were recently cloned and characterized. Yac-1 enzyme possesses ADP-ribosyltransferases activity. In contrast, Yac-2 has significant NAD glycohydrolase activity and may preferentially hydrolyze NAD. Yac-2 possesses a glutamate at position 219 adjacent to the two consdrved glutamic acid residues. To study the effect of Glu-219 on enzyme activities, Glu-219 was mutagenized to Glutamine (E219Q) or alanine (E219A) using a two-step recombinant polymerase chain reaction procedure. Replacing Glu at position 219 with Gln or Ala resulted in 56 (E219Q) or 66% (E219A) reduction in ADP-ribosyltranferase activity. The NAD glycohydrolase activity of Yac-2 protein were not altered by the mutations. These results indicate that Glu-219 in Yac-2 enzyme plays an important role in ADP-ribosyltransferase, but not NAD glycohydrolase activity.

두 종류의 ADP-riboyltransferase(Yac-1과Yac-2)가 최근 mouse의 임파구로부터 cloning되어 그 특성이 규명되어졌다. Yac-1효소는 ADP-ribosyltransferase활성을 보여주나, 대조적으로, Yac-2는 상당한 양의 NAD glycohydrolase 활성도 소유하고` 있으며 이는 NDA를 우선적으로 가수분해 할 수 있다는 사실을 반영한다. Yac-2는 보존된 두 glutamate에 인접한 위치인 219번에 또 다른 glutamate를 소유하고 있다. 효소 활성에 대한 Glu-219의 효과를 알아보기 위해 두 단계의 재조합 중합효소 연쇄 반응 방법에 의해 Glu-219가 glutamate (E219Q)혹은 alanine(E219A)으로 치환되었다. Gln혹은 Ala으로의 치환결과, ADP-ribosyltransferase활성은 56% (E219Q)혹은 66% *E219A)로 감소하였다. Yac-2단백질의 NAD glycohydrolase 활성은 돌연변이에 의해 영향을 받지 않았다. 이러한 결과는 Yac-2효소의 Glu-219가 ADP-ribosyltransferase 활성에 중요한 역할을 하나, NDA glycohydrolase활성에는 관여하지 않음을 시사한다.

Keywords

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