Disulfide bonds and the translocation of proteins across membranes.
We are concerned with the mechanisms whereby hydrophilic proteins synthesized in the cytoplasm are translocated across one or two membranes into different cellular organelles. On the basis of a model of the translocation process to be described elsewhere, we propose an explanation of previous findings that the in vitro translocation across the endoplasmic reticulum of secretory proteins of higher eukaryotic cells appears to be obligatorily co-translational (i.e., occurs only while the polypeptide chain is being synthesized on the ribosome). We suggest that in vitro the intrachain disulfide bonds of the polypeptide rapidly form after it is released from the ribosome; the three-dimensional conformation of the chain is thereby stabilized and cannot undergo the unfolding that is required for post-translational translocation. In accord with this proposal, we show that the secretory preprotein human preprolactin, after translation and release from the ribosome, can indeed undergo translocation across endoplasmic reticulum membranes in vitro if the medium is sufficiently reducing. Those polypeptides that, in the absence of reducing agents, can be post-translationally translocated in vitro across bacterial, mitochondrial, and other types of membranes may generally lack intrachain disulfide bonds.
Maher
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PA
Singer
S J
SJ
D001692:Biological Transport / Q000187:drug effects; D003545:Cysteine / Q000502:physiology; D004220:Disulfides / Q000502:physiology; D004229:Dithiothreitol / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000378:metabolism; D066298:In Vitro Techniques; D008565:Membrane Proteins / Q000378:metabolism; D008861:Microsomes / Q000378:metabolism; D011388:Prolactin / Q000378:metabolism; D011487:Protein Conformation; D011498:Protein Precursors / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000378:metabolism*
D004220:Disulfides; D008565:Membrane Proteins; D011498:Protein Precursors; D011506:Proteins; C017671:preprolactin; D011388:Prolactin; D003545:Cysteine; D004229:Dithiothreitol