ヒト前立腺アンドロゲン・レセプター精製に関する研究 第1編: affnity chromatograhpyを使用した精製法

Abstract

前立腺肥大症の腺腫組織1, 700 gよりaffinity chromatographyを用いてアンドロゲン・レセプター(AR)の精製を試みた.1)最終精製分画は3H-R1881と結合能を有し, その結合は1, 000倍量のtriamcinolone acetonide (TA)でも抑制されなかった.2) Scatchard法による検討では最終精製分画の解離定数は2.3 nMと高親和性の結合能を有し, ARの結合特異性を残していた.3)精製倍率は蛋白量あたりの結合能で計算すると約38倍であった.4) Polyacrylamide gel electrophoresis (PAGE)による検討では, albuminやtestosterone-estradiol binding globulin (TeBG)の混入を認めなかった

The androgen receptor (AR) was purified to establish a new method for measuring AR after producing the antibody for the purified AR. AR was purified from 1, 700 g of benign prostatic hypertrophic tissue by combining affinity chromatography of heparin Sepharose CL-6B and of 17 alpha-carboxy-hexamethyl-17-hydroxy-4-androstane-3-one Sepharose 4B. The final purification fraction had the high affinity, low capacity binding to 3H-R 1881 in the presence of 1, 000-fold molar excess of triamcinolone acetonide with a dissociation constant of 2.3 nM, showing the binding specificity of AR. Polyacrylamide gel electrophoresis did not reveal any albumin or TeBG mixed in the final purification fraction. These results showed that AR was partially purified, even though the degree of purification was low, around 38 fold calculated by the binding capacity per mg protein.