Transmembrane biogenesis of Kv1.3

Liwei Tu, Jing Wang, Andrew Helm, William Skach, Carol Deutsch

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

Using a combination of protease protection, glycosylation, and carbonate extraction assays, we have characterized the topogenic determinants encoded by Kv1.3 segments that mediate translocation events during endoplasmic reticulum (ER) biogenesis. Transmembrane segments S1, S2, S3, S5, and S6 initiate translocation, only S1 and S2 strongly (>60%) anchor themselves in the membrane, S5 exhibits signal anchor activity and contains a cryptic cleavage site, and S3 and S6 fail to integrate into the membrane. Elongation of each single-transmembrane construct to include multiple transmembrane segments alters integration and translocation efficiencies, indicating that multiple topogenic determinants cooperate during Kv1.3 topogenesis and assembly. Several surprising findings emerged from these studies. First, in the presence of T1, the N-terminal recognition domain, S1 was unable to initiate either translocation or membrane integration. As a result, S2 likely functions as the initial signal sequence to establish Kv1.3 N-terminus topology. Second, S4 independently integrates into the membrane. Third, S6 plus the C-terminus of Kv1.3 is a secretory protein but can be converted to a membrane-integrated protein with a correctly oriented, cytosolic C-terminus by linking S6 to S5 and the pore loop. These results have implications for the role of the N-terminus in Kv biogenesis and on the mechanisms of dominant negative suppression of Kv1.3 by truncated Kv1.3 fragments [Tu et al. (1996) J. Biol. Chem. 271, 18904-18911].

Original languageEnglish (US)
Pages (from-to)824-836
Number of pages13
JournalBiochemistry
Volume39
Issue number4
DOIs
StatePublished - Feb 1 2000

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S 6
Membranes
Anchors
Glycosylation
Carbonates
Protein Sorting Signals
Endoplasmic Reticulum
Elongation
Assays
Membrane Proteins
Peptide Hydrolases
Topology
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Tu, L., Wang, J., Helm, A., Skach, W., & Deutsch, C. (2000). Transmembrane biogenesis of Kv1.3. Biochemistry, 39(4), 824-836. https://doi.org/10.1021/bi991740r

Transmembrane biogenesis of Kv1.3. / Tu, Liwei; Wang, Jing; Helm, Andrew; Skach, William; Deutsch, Carol.

In: Biochemistry, Vol. 39, No. 4, 01.02.2000, p. 824-836.

Research output: Contribution to journalArticle

Tu, L, Wang, J, Helm, A, Skach, W & Deutsch, C 2000, 'Transmembrane biogenesis of Kv1.3', Biochemistry, vol. 39, no. 4, pp. 824-836. https://doi.org/10.1021/bi991740r
Tu L, Wang J, Helm A, Skach W, Deutsch C. Transmembrane biogenesis of Kv1.3. Biochemistry. 2000 Feb 1;39(4):824-836. https://doi.org/10.1021/bi991740r
Tu, Liwei ; Wang, Jing ; Helm, Andrew ; Skach, William ; Deutsch, Carol. / Transmembrane biogenesis of Kv1.3. In: Biochemistry. 2000 ; Vol. 39, No. 4. pp. 824-836.
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