Role of ATP and disulphide bonds during protein folding in the endoplasmic reticulum.
Being topologically equivalent to the extracellular space, the lumen of the endoplasmic reticulum (ER) provides a unique folding environment for newly synthesized proteins. Unlike other compartments in the cell where folding occurs, the ER is oxidizing and therefore can promote the formation of disulphide bonds. The reducing agent dithiothreitol, when added to living cells, inhibits disulphide formation with profound effects on folding. Taking advantage of this effect, we demonstrate here that folding of influenza haemagglutinin is energy dependent. Metabolic energy is required to support the correct folding and disulphide bond formation in this well characterized viral glycoprotein, to rescue misfolded proteins from disulphide-linked aggregates, and to maintain the oxidized protein in its folded and oligomerization-competent state.
Braakman
I
I
Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06510-8002.
Helenius
J
J
Helenius
A
A
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D004220:Disulfides / Q000378:metabolism*; D004229:Dithiothreitol / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D004734:Energy Metabolism; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral / Q000096:biosynthesis* / Q000737:chemistry*; D011487:Protein Conformation / Q000187:drug effects
D004220:Disulfides; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral; D000255:Adenosine Triphosphate; D004229:Dithiothreitol