%0 Journal Article
%T Reversed-phase high performance liquid chromatographic behavior of unfolding procedure of Huwentoxin-V]<br>虎纹捕鸟蛛毒素-Ⅴ去折叠过程的色谱分析
%A ZHANG Pengfei~
%A <br>张鹏飞
%A 肖顺勇
%A 梁宋平
%J 色谱
%D 2004
%I 
%X Huwentoxin-V (HWTX-V) is an insecticidal peptide purified from the venom of spider Selnocosmia huwena. HWTX-V contains 33 amino acid residues, including six cysteine residues that form three pairs of disulfide bond. A method for measuring unfolding intermediates of HWTX-V by reversed-phase high performance liquid chromatography (HPLC) has been established. HWTX-V was first denatured over 30 min in the guanidine hydrochloride, and then reduced by using tris-(2-carboxyethyl)-phosphine at pH 3.0 over 12 min. All intermediates were separated on a C18 column (4.6 mm i. d. x 390 mm) with a linear gradient elution of acetonitrile containing 0.1% (v/v) trifluoroacetic acid, and identified by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS). Finally, these intermediates were carboxamidomethylated by iodoacetamide at the concentration of 0.5 mol/L and verified by MALDI-TOF MS. The diverse chromatographic retention behaviors for intermediates of Huwentoxin-V are discussed. This method is helpful to reveal the conformation changes in the procedures of proteins/peptides unfolding.
%K NULL<br>反相高效液相色谱
%K 基体辅助激光解吸电离-飞行时间质谱
%K 三羧甲基磷酸
%K 去折叠
%K 虎纹捕鸟蛛毒素-V
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=6E709DC38FA1D09A4B578DD0906875B5B44D4D294832BB8E&cid=6579068328FE643F&jid=4D81E042D77AFEC6881D14759692069C&aid=3134A9B40C7DBD88&yid=D0E58B75BFD8E51C&vid=BC12EA701C895178&iid=E158A972A605785F&sid=A22854835F81B3F8&eid=23F20F9780C3579E&journal_id=1000-8713&journal_name=色谱&referenced_num=0&reference_num=8