Different conformations of nascent polypeptides during translocation across the ER membrane.
BACKGROUND
In eukaryotic cells, proteins are translocated across the ER membrane through a continuous ribosome-translocon channel. It is unclear to what extent proteins can fold already within the ribosome-translocon channel, and previous studies suggest that only a limited degree of folding (such as the formation of isolated alpha-helices) may be possible within the ribosome.


RESULTS
We have previously shown that the conformation of nascent polypeptide chains in transit through the ribosome-translocon complex can be probed by measuring the number of residues required to span the distance between the ribosomal P-site and the lumenally disposed active site of the oligosaccharyl transferase enzyme (J. Biol. Chem 271: 6241-6244). Using this approach, we now show that model segments composed of residues with strong helix-forming properties in water (Ala, Leu) have a more compact conformation in the ribosome-translocon channel than model segments composed of residues with weak helix-forming potential (Val, Pro).


CONCLUSIONS
The main conclusions from the work reported here are (i) that the propensity to form an extended or more compact (possibly alpha-helical) conformation in the ribosome-translocon channel does not depend on whether or not the model segment has stop-transfer function, but rather seems to reflect the helical propensities of the amino acids as measured in an aqueous environment, and (ii) that stop-transfer sequences may adopt a helical structure and integrate into the ER membrane at different times relative to the time of glycan addition to nearby upstream glycosylation acceptor sites.
Mingarro
I
I
Department of Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden. ismael.mingarro@uv.es
Nilsson
I
I
Whitley
P
P
von Heijne
G
G
D000409:Alanine / Q000737:chemistry; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D004285:Dogs; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D006031:Glycosylation; D007425:Intracellular Membranes / Q000378:metabolism*; D007930:Leucine / Q000737:chemistry; D026901:Membrane Transport Proteins / Q000737:chemistry; D008861:Microsomes / Q000378:metabolism; D008969:Molecular Sequence Data; D010449:Peptide Mapping / Q000379:methods; D011392:Proline / Q000737:chemistry; D011487:Protein Conformation; D021381:Protein Transport; D012270:Ribosomes / Q000737:chemistry; D018271:Signal Recognition Particle / Q000378:metabolism*; D014633:Valine / Q000737:chemistry
D026901:Membrane Transport Proteins; D018271:Signal Recognition Particle; D011392:Proline; D007930:Leucine; D014633:Valine; D000409:Alanine