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摘 要：利用反相高效液相色谱（RP-HPLC）和离子交换色谱（IEC）比较研究了固相化学合成的多肽类神经毒素虎纹捕鸟蛛毒素-Ⅰ（HWTX-Ⅰ）与其对应的天然产物的色谱行为差异及分离纯化方法。发现在完全相同的实验条件下，复性处理后的合成HWTX-Ⅰ的谱峰有别于还原变性后又复性的天然HWTX-Ⅰ的谱峰，主要表现为扩散度较大，对称性较差，说明合成产物存在更大分子构象的不均一性。推测除了在合成HWTX-Ⅰ复性过程中有少数分子发生二硫键错配外，还有部分分子在固相合成中发生了消旋作用，需要交替采取多种色谱法才能完全分离纯化复性后的合成多肽产物。
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Abstract：A
comparative study was made to investigate the chromatographic behavior
and purification of chemically synthesized and native polypeptide
neurotoxin huwentoxin-Ⅰ(HWTX-Ⅰ)
by means of reversed-phase HPLC and ion-exchange chromatography. The
results showed that the synthetic HWTX-Ⅰ in
crude product has a longer retention time than the native HWTX-Ⅰ,but
has the same retention time as the denatured HWTX-Ⅰ,
suggesting it being in a state of complete denaturation which was
confirmed by the MALDI-TOF MS analysis. It has also been discovered
that, before complete purification, the synthetic HWTX-Ⅰ,
after oxidative treatment, showed wider and more asymmetric peaks than
the native HWTX-Ⅰ that
was denatured and then renatured under the same experimental conditions,
which suggests that there is even greater conformational unhomogeneity
in the synthetic HWTX-Ⅰ.
It was inferred that, in addition to the wrong disulfide bonds formation
during oxidative folding, the racemization during chemical synthesis
resulted in more stereoisomers of synthetic HWTX-Ⅰ.
It was needed for the renatured synthetic peptides to be purified using
different methods alternatively. |
| 基金项目：国家863高技术发展计划资助(103-13-01-06) 作者简介：王贤纯(1959-)，男，博士，教授。 作者单位：王贤纯（湖南师范大学生命科学学院，湖南长沙 410081） 梁宋平（湖南师范大学生命科学学院，湖南长沙 410081） 参考文献： ［1］Zhang D Y, Liang S P. J Protein Chem,
1993,12(6):735-740 |
原载于《色谱》2001 Vol.19 No.2
P.101-104
http://periodical.wanfangdata.com.cn/periodical/sp/sp2001/0101/0101ml.htm