Expression, Purification and Characterization of Gly14-Humanin, a Novel Neuroprotective Peptide

VernacularTitle:新型神经保护肽[Gly14]-Humanin的表达,纯化和活性鉴定
Author: Bao-Feng YU ¹ ; Jun XIE ; Xian-Jiu CHEN ; Yue-Hong ZHANG ; Hui-Zhen WANG ; Niu-Liang CHENG ; Bo NIU
Author Information

1. 山西医科大学
Keywords: Neuroprotection [Gly14]-Humanin (HNG) Expression
From: China Biotechnology 2008;28(4):21-26
CountryChina
Language:Chinese
Abstract: Humanin (HN, its analogue [Gly14]-Humanin, HNG) was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults. But the relative low content of this peptide in its natural sources limits its further characterization. An expression vector pET32a/HNG was corstructed and transformed it into E. coli BL21 trxB (DE3). HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography. Subsequently, the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC. A 23 mg recombinant HNG (rHNG) from 1 L bacterial culture was purified. The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide. The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence. Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.