The CXXCXXC motif determines the folding, structure and stability of human Ero1-Lalpha.
The presence of correctly formed disulfide bonds is crucial to the structure and function of proteins that are synthesized in the endoplasmic reticulum (ER). Disulfide bond formation occurs in the ER owing to the presence of several specialized catalysts and a suitable redox potential. Work in yeast has indicated that the ER resident glycoprotein Ero1p provides oxidizing equivalents to newly synthesized proteins via protein disulfide isomerase (PDI). Here we show that Ero1-Lalpha, the human homolog of Ero1p, exists as a collection of oxidized and reduced forms and covalently binds PDI. We analyzed Ero1-Lalpha cysteine mutants in the presumed active site C(391)VGCFKC(397). Our results demonstrate that this motif is important for protein folding, structural integrity, protein half-life and the stability of the Ero1-Lalpha-PDI complex.
Benham
A M
AM
Department of Biochemistry, Academic Medical Center, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands.
Cabibbo
A
A
Fassio
A
A
Bulleid
N
N
Sitia
R
R
Braakman
I
I
D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D004220:Disulfides / Q000737:chemistry; D004721:Endoplasmic Reticulum / Q000737:chemistry; D006031:Glycosylation; D015699:HIV Envelope Protein gp120 / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D008562:Membrane Glycoproteins / Q000737:chemistry*; D010084:Oxidation-Reduction; D010088:Oxidoreductases; D011487:Protein Conformation; D017510:Protein Folding
D004220:Disulfides; D015699:HIV Envelope Protein gp120; D008562:Membrane Glycoproteins; C405327:ERO1A protein, human; D010088:Oxidoreductases