Abstract
Human Bence Jones Protein could be purified by DEAF-Sephadex A-50 column $(2{\times}37cm)$ with 0.02M phosphate Buffer (pH 8.0) and gradient increasing with NaCl concentration as in Fig. 2-4. Sample As (K-type Bence Jones Protein) had two component, F-I was major component and its dried weight was 350mg. of starting material of 500mg. Other Sample Im and Ik (${\lambda}$-type Bence Jones Protein) was purified by DEAE-Sephadex A-50 with 0.02M phosphate Buffer(pH 8.0)too. F-I (major component) of Im and F-I of Ik were 242mg and 146mg. its dried weight respectively. K-type of Bence Jones Protein's(As, Ko, Ta.) N-terminal amino acid residue was determined by method of DNP,. K-type of Bence Jones Protein's amino acid residue were either glutamic acid or aspartic acid. Sample Ta was confirmed as glutamic acid its N-Terminal. As and Ko were aspartic acid. Each yellowish spot (DNP-amino acids) were extracted with 4ml. of pH 8.05% $NaHCO_3$ solution and calculated its recovery by O.D. $(360m{\mu}$ using the ${\varepsilon}=18.1{\times}10^3DNP$ $Asp\;{\varepsilon}=17.41{\times}10^(3)\;DNP\;Glu$ considering 50% lose during; the acid (6N-HCI) hydrolysis. Recovery of ko and As were 54.3% and 65% of its starting materials (DNP-Protein). Sample Ta's recovery was 85% of its DNP-protein. ${\lambda}$-type of Bence Jones Protein was rot investigated its N-terminal amino acid residue by DNP-method, probably it was blocked its N-terminal residue with glutamic acid.
Bence Jones 단백질(蛋白質)(As,Ik,In)를 DEAE-Sephadex A-50 Column으로 0.02M phosphate Buffer(pH8.)와 0.5M NaCl를 gradient로 사용(使用)해 정재(精製)하였다. 시료(試料) As(K-형(型))의 정재결과(精製結果) 주성분(主成分)인 F-1는 조단백질(粗蛋白質) 500mg로부터 350mg 얻었으며 다른 시료(試料) Im와 Ik($lambda$-형(型))에서도 각각 주성분(主成分)을 242mg(Im)와 146mg(Ik) 얻을 수 있었다. 정재(精製)한 시료(試料)는 그의 순도(純度)를 알기 위해 Poly acryl amide의 Disc electrophoresis로 확인한 후 K-형(型) Bence jones단백질(蛋白質)의 일종(一種)인 As와 $lambda$-형(型)의 일종(一種)인 IM를 6N-HCI로 $105^{\circ}C$에서 20시간(時間) 가수분해(加水分努)후 Amino 산(酸) 조정(組成)을 살펴보았다. DNP법(法)으로 시료(試料) K-형(型)(As, Ko, Ta)의 단백질(蛋白質)의 N-말단(末端)을 검출(檢出)해 Ta는 glutamic acid, Ko, As는 Aspartic acid임을 확인하였으며 Paper 상(上)에 나타난 DNP-Amino 산(酸)은 5% $NaHCO_3$ (pH8) 4ml에 추출(抽出)해 그 수량(收量)을 ${\varepsilon}=18.1{\times}10^3DNP$ $Asp\;{\varepsilon}=17.41{\times}10^(3){\;}DNP{\;}Glu$에 의(依)해 산출(算出)하였더니 Ko는 54.3%, As는 65%이였으며 Ta는 85%의 수량(收量)이였다. ${\lambda}$-형(型)의 Im,Ik의 시료(試料)도 DNP법(法)으로 N-말단(末端)을 검출(檢出)해 보았더니 검출(檢出)되지 아니하였다.