Transmembrane Segments Form Tertiary Hairpins in the Folding Vestibule of the Ribosome

Overview

Journal J Mol Biol

Publisher Elsevier

Specialties Microbiology
Molecular Biology

Date 2013 Sep 24

PMID 24055377

Citations 39

Authors

LiWei Tu

Pooja Khanna

Carol Deutsch

Affiliations

Soon will be listed here.

Abstract

Folding of membrane proteins begins in the ribosome as the peptide is elongated. During this process, the nascent peptide navigates along 100Å of tunnel from the peptidyltransferase center to the exit port. Proximal to the exit port is a "folding vestibule" that permits the nascent peptide to compact and explore conformational space for potential tertiary folding partners. The latter occurs for cytosolic subdomains but has not yet been shown for transmembrane segments. We now demonstrate, using an accessibility assay and an improved intramolecular crosslinking assay, that the helical transmembrane S3b-S4 hairpin ("paddle") of a voltage-gated potassium (Kv) channel, a critical region of the Kv voltage sensor, forms in the vestibule. S3-S4 hairpin interactions are detected at an early stage of Kv biogenesis. Moreover, this vestibule hairpin is consistent with a closed-state conformation of the Kv channel in the plasma membrane.

Citing Articles

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The ribosome stabilizes partially folded intermediates of a nascent multi-domain protein.

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References

Alabi A, Bahamonde M, Jung H, Kim J, Swartz K . Portability of paddle motif function and pharmacology in voltage sensors. Nature. 2007; 450(7168):370-5. PMC: 2709416. DOI: 10.1038/nature06266. View

Lu J, Hua Z, Kobertz W, Deutsch C . Nascent peptide side chains induce rearrangements in distinct locations of the ribosomal tunnel. J Mol Biol. 2011; 411(2):499-510. PMC: 3143575. DOI: 10.1016/j.jmb.2011.05.038. View

Hardesty B, Kramer G . Folding of a nascent peptide on the ribosome. Prog Nucleic Acid Res Mol Biol. 2000; 66:41-66. DOI: 10.1016/s0079-6603(00)66026-9. View

Long S, Campbell E, MacKinnon R . Voltage sensor of Kv1.2: structural basis of electromechanical coupling. Science. 2005; 309(5736):903-8. DOI: 10.1126/science.1116270. View

OBrien E, Hsu S, Christodoulou J, Vendruscolo M, Dobson C . Transient tertiary structure formation within the ribosome exit port. J Am Chem Soc. 2010; 132(47):16928-37. DOI: 10.1021/ja106530y. View

Broomand A, Elinder F . Large-scale movement within the voltage-sensor paddle of a potassium channel-support for a helical-screw motion. Neuron. 2008; 59(5):770-7. DOI: 10.1016/j.neuron.2008.07.008. View

Jiang Y, Lee A, Chen J, Ruta V, Cadene M, Chait B . X-ray structure of a voltage-dependent K+ channel. Nature. 2003; 423(6935):33-41. DOI: 10.1038/nature01580. View

Mingarro I, Nilsson I, Whitley P, von Heijne G . Different conformations of nascent polypeptides during translocation across the ER membrane. BMC Cell Biol. 2001; 1:3. PMC: 29064. DOI: 10.1186/1471-2121-1-3. View

Long S, Tao X, Campbell E, MacKinnon R . Atomic structure of a voltage-dependent K+ channel in a lipid membrane-like environment. Nature. 2007; 450(7168):376-82. DOI: 10.1038/nature06265. View

10.

Lu J, Deutsch C . Secondary structure formation of a transmembrane segment in Kv channels. Biochemistry. 2005; 44(23):8230-43. DOI: 10.1021/bi050372q. View

11.

Huang F, Chen J, Lin M, Keating M, Sanguinetti M . Long-QT syndrome-associated missense mutations in the pore helix of the HERG potassium channel. Circulation. 2001; 104(9):1071-5. DOI: 10.1161/hc3501.093815. View

12.

Hou B, Lin P, Johnson A . Membrane protein TM segments are retained at the translocon during integration until the nascent chain cues FRET-detected release into bulk lipid. Mol Cell. 2012; 48(3):398-408. PMC: 3496027. DOI: 10.1016/j.molcel.2012.08.023. View

13.

Borel A, Simon S . Biogenesis of polytopic membrane proteins: membrane segments of P-glycoprotein sequentially translocate to span the ER membrane. Biochemistry. 1996; 35(33):10587-94. DOI: 10.1021/bi960950q. View

14.

Egea P, Stroud R . Lateral opening of a translocon upon entry of protein suggests the mechanism of insertion into membranes. Proc Natl Acad Sci U S A. 2010; 107(40):17182-7. PMC: 2951439. DOI: 10.1073/pnas.1012556107. View

15.

Hessa T, White S, von Heijne G . Membrane insertion of a potassium-channel voltage sensor. Science. 2005; 307(5714):1427. DOI: 10.1126/science.1109176. View

16.

Kida Y, Morimoto F, Sakaguchi M . Two translocating hydrophilic segments of a nascent chain span the ER membrane during multispanning protein topogenesis. J Cell Biol. 2008; 179(7):1441-52. PMC: 2373506. DOI: 10.1083/jcb.200707050. View

17.

Lu J, Kobertz W, Deutsch C . Mapping the electrostatic potential within the ribosomal exit tunnel. J Mol Biol. 2007; 371(5):1378-91. DOI: 10.1016/j.jmb.2007.06.038. View

18.

Cross B, High S . Dissecting the physiological role of selective transmembrane-segment retention at the ER translocon. J Cell Sci. 2009; 122(Pt 11):1768-77. DOI: 10.1242/jcs.046094. View

19.

Kowarik M, Kung S, Martoglio B, Helenius A . Protein folding during cotranslational translocation in the endoplasmic reticulum. Mol Cell. 2002; 10(4):769-78. DOI: 10.1016/s1097-2765(02)00685-8. View

20.

Bhushan S, Gartmann M, Halic M, Armache J, Jarasch A, Mielke T . alpha-Helical nascent polypeptide chains visualized within distinct regions of the ribosomal exit tunnel. Nat Struct Mol Biol. 2010; 17(3):313-7. DOI: 10.1038/nsmb.1756. View