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An efficient and reliable electroelution method from SDS-PAGE: Identification of a 31 kDa protein in the postsynaptic density fraction as adenine nucleotide translocator 1

SDS-겔로부터 효율적인 단백질 분리방법과 31 kDa 연접후치밀질 단백질의 동정

  • Jung, Jae-Seob (Department of Microbiology, College of Natural Sciences, Kyungpook National University) ;
  • Cho, Sun-Jung (Department of Microbiology, College of Natural Sciences, Kyungpook National University) ;
  • Shin, Seung-Chul (Department of Anatomy and Research institute for Natural Science of Dongguk University) ;
  • Jin, Ing-Nyol (Department of Microbiology, College of Natural Sciences, Kyungpook National University) ;
  • Jung, Yong-Wook (Department of Anatomy and Research institute for Natural Science of Dongguk University) ;
  • Ko, Bok-Hyun (Department of Anatomy and Research institute for Natural Science of Dongguk University) ;
  • Moon, Il-Soo (Department of Anatomy and Research institute for Natural Science of Dongguk University)
  • Published : 2002.10.01

Abstract

The molecular composition of the postsynaptic density (PSD) is largely hon. In this report, an electroelution protocol was demonstrated to be used for efficient isolation of PSD proteins with diverse molecular sizes. Using this protocol, a 31 kDa protein in the 1% n-octyl glucoside-insoluble PSD fraction (termed as PSD31) was purified from SDS-gels, and internal peptides were determined for amino acid sequences. The amino acid sequences of the PSD31 were highly homologous with the adenine nucleotide translocator 1 (ANTI). The association of ANTl with PSD suggests presence of a mechanism in synapses for releasing adenosine nucleotides into the extracellular space.

연접후치밀질(PSD)의 분자조성은 아직 대체로 알려져 있지 않다. 본 연구에서는 다양한 크기의 연접후치밀질 단백질들을 효율적으로 SDS-겔에서부터 추출하는 방법을 제시하고, 이를 이용하여 연접후치밀질의 1% n-octyl glucoside에 용해되지 않는 분핵에 존재하는 31kDa크기의 단백직(PSD31로 명명)을 SDS-겔로부터 분리하고, trypsin 처리하여 생성되는 펩티드의 아미노산 서열을 결정하였다. PSD31의 아미노산 서열은 adenine nucleotide translocator 1(ANT1)과 매우 상동성이 높았다. 연접후치밀질에 ANT1이 존재함은 신경조직의 세포외 공간에 adenosine nucleotide를 유리할 수 있는 기구가 연접에 존재함을 시사한다.

Keywords

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