Purification and structural characterization of human ERp29.
ERp29 is a major resident of the endoplasmic reticulum (ER) and is postulated to play an important molecular chaperone role in most animal cells. Human ERp29 was isolated to homogeneity in high yield by using a bacterial expression system. Its secondary structure was studied by circular dichroism (CD), Fourier transformed infrared spectroscopy (FTIR) and Raman spectroscopy and it was found that human ERp29 comprises significant alpha-helical structure. The details of its temperature-induced conformational changes was studied by CD and FTIR for the first time, revealing that the protein is stable below 50 degrees C and has two distinct structural transitions between 50 degrees C and 70 degrees C. This may shed light on ERp29's inability to protect substrate proteins against thermal aggregation.
Zheng
Jinbiao
J
Center of Analysis and Measurement, Fudan University, Shanghai 200433, PR China.
Liu
Xingang
X
Yan
Xiaomin
X
Dai
Linsen
L
Ji
Chaoneng
C
D000595:Amino Acid Sequence; D000818:Animals; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D017550:Spectroscopy, Fourier Transform Infrared; D013059:Spectrum Analysis, Raman
C491738:ERP29 protein, human; C110909:Erp29 protein, rat; D006360:Heat-Shock Proteins; D011994:Recombinant Proteins