글루코즈-6-인산 가수분해 효소에 $^{32}P$로 표시된 글루코즈-6-인산을 과량 넣어 주어서 steady-state의 상태에서 글루코즈-6-인산 가수분해 효소를 표지시킨다. 쥐간에서 초고속 원심분리 방법으로 마이크로좀을 얻고 이 단백질 혼합물에$[^{32}P]$ 글루코즈-6-인산을 넣어 반응 후, SDS 젤에서 전기영동으로 분리, 건조하여 X-ray film으로 감광시킨 결과 60 kd의 한 밴드를 보았다. 한편, 만노즈-6-인산과 갈락토즈-6-인산을 기질로 사용해서도 똑같은 밴드를 보았다. 또한 단백질 변성 방법으로 8M 우레아를 쓰거나, 20% Trichloro-acetic acid 혹은 2% SDS를 사용해도 여전히 같은 위치에서 밴드가 나타난다.
Glucose-6-phosphatase has been known as a multicomponent system and glucose-6-phosphatase was labeledd by incubating with $[^{32}P]$glucose-6-phosphate to be saturation concentration. Microsomal pellet was obtained by the method of differential ultracentrifugation and distrupted with sonication. To this microsomal protein mixture, $[^{32}P]$glucose-6-phosphate was added and incubated. Then the mixture was seperated with 10∼15% gradient PAGE and the gel was dried and exposed on the X-ray film. As a result, 60 kd band was obtained and this was considered as a glucose-6-phosphatase. The same result was obtained when $[^{32}P]$mannose-6-phosphate and $[^{32}P]$galactose-6-phosphate were used as substrates. 20% trichloro acetic acid. 8 M urea, and 10% SDS were used as a quenching system respectively and 60 kd band was still unchanged.
