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標題: | 豬腦下腺生長激素之分離與純化及其刺激肝內烏胺酸去羧基?之作用 ISOLATION AND CEARACTERIZATION OF PORCINE GROWTH HORMONE AND ITS INDUCTION OF HEPATIC ORNITHINE DECARBOXYLASE |
作者: | Jiann-Jau Jane 簡健釗 |
出版年 : | 1982 |
學位: | 碩士 |
摘要: | 生長激素為腦下腺前葉所分泌之聚合勝?。目前確知有促進骨骼與內臟生長之作用。並有促進脂解,造成氨基酸聚集之作用。其中尤要□乃促進蛋白質之合成。 本研究乃探究豬生長激素之分離、純化之最佳方法。並研究其是否□人生長激素般促進烏胺酸去羧基?(ornithine decarboxylase)之作用。(該?促進多胺類合成而影響DNA合成、進而影響蛋白質之合成。) 實驗結果發現,依照人生長激素之分離方法可得粗的豬生長激素,進一步之純化方法?: (一)先用Gel-Filtration法。 (Sephacryl S-200,緩衝溶液用0.05N之NH4HCO3,PH8.0左右。) (二)再用Ion-Exchange Chromatography。 (DEAE-sephadex A-25,緩衝溶液仍用NH4HCO3,濃度變化?0.01N,0.2N,0.4N,1.0N,逐步加入。) 分析發現,其分子量約?22,000,N-端?苯氨基丙酸(phenyl-alanine)。α-螺旋約66.7%。極易聚合成多聚物而沉澱。此外尚□了氨基酸組成,電泳,CD, HPLC等分析。 生物分析方面、發現確能促進小白鼠之生長。 在對烏胺酸去羧基?之促進實驗中,發現隨著純化的程度,促進該?活性之能力隨增。故此種實驗方法似可做?生長激素純度之指標。有最高作用之時間?注射生長激素後四小時;且會做牛生長激素、牛催乳激素等之□與其比較。 純化後所取之四不同成分均有促進該?活性之作用。故懷疑此些皆?生長激素,只是結構上有變異。(如同人生長激素一樣。)此尚待進一步之□。 This report describes a method for the isolation and purification of porcine growth hormone (PGH) from the pituitary glands, as shown in the following: 1. Extraction of PGH at PH 10.5 followed by fractional precipctation at 0.2-0.4 saturation of ammonium sulfate. 2. Gel-filtration on Sephacryl S-200 column. 3. Ion-Exchange chromatography on DEAE-Sephadex A-25, eluted with NH4HCO3 solution. The molecular weight was about 22,000 the N-terminal residue was phenylalanine. The α-helix content was about 66.7%, It was very easy to aggregate as dimer or polymers. In addition, we also analyzed its amino acid composition, homogeneity and HPLC patterns. We found that it indeed promoted the growth of mice. We found that as the purity of PGH increased, the ability to induce the hepatic ODC activity increased. This could be used to monitor the purity of PGH. also, the maximum induction time was 4 hours after the injection of PGH. For comparison, we also tested BGH and BPRL. The four fractions from DEAE-Sephadex A-25 colutnn were able to induce hepatic ODC. We suspected that these were all the PGH differing from one another in some aspects. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75417 |
全文授權: | 未授權 |
顯示於系所單位: | 生化科學研究所 |
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