[Chromatographic behavior and purification of synthetic huwentoxin-I].
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.
Wang
X C
XC
College of Life Science, Hunan Normal University, Changsha 410081, China.
Liang
S P
SP
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange; D009498:Neurotoxins / Q000138:chemical synthesis / Q000302:isolation & purification*; D030162:Reptilian Proteins; D013111:Spider Venoms / Q000138:chemical synthesis / Q000302:isolation & purification*
D009498:Neurotoxins; D030162:Reptilian Proteins; D013111:Spider Venoms; C083624:huwentoxin I