Evidence for interaction between transmembrane segments in assembly of Kv1.3

ZuFang Sheng, William Skach, Vincent Santarelli, Carol Deutsch

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Previously, we showed that the N-terminal recognition domain (T1) of Kv1.3 was not required for assembly of functional channels [Tu et al. (1996) J. Biol. Chem. 271, 18904-18911]. Moreover, specific Kv1.3 peptide fragments including regions of the central core are able to inhibit expression of current produced from a channel lacking the T1 domain, Kv1.3(T1-). To elucidate the mechanism whereby Kv1.3 peptide fragments suppress Kv1.3(T1-) current, we have studied the ability of peptide fragments containing the transmembrane segments S1, S1-S2, or S1-S2-S3 to physically associate with the Kv1.3(T1-) polypeptide subunit in vitro in microsomal membranes. Using c-myc (9E10) epitope-labeled peptide fragments and anti-myc antibody as well as antisera to the Kv1.3 C-terminus, we now demonstrate specific association of these peptide fragments with Kv1.3(T1-). Association of peptide fragments with Kv1.3(T1-) was correlated with integration of both proteins into the membrane. Furthermore, the relative strength and kinetics of this association directly correlated with the ability of fragments to suppress Kv1.3(T1-) current. The rate-limiting step in the sequential synthesis, integration, and formation of a complex was the association of integrated polypeptides within the plane of the lipid bilayer. These results strongly suggest that the physical association of transmembrane segments provides the basis for suppression of K+ channel function by K+ channel peptide fragments in vivo. Moreover, the S1-S2-S3 peptide fragment potently suppressed full-length Kv1.3, thus implicating a role for the S1-S2-S3 region of Kv1.3 in the assembly of the Kv1.3 channel. We refer to these putative association sites as IMA (intramembrane association) sites.

Original languageEnglish (US)
Pages (from-to)15501-15513
Number of pages13
JournalBiochemistry
Volume36
Issue number49
DOIs
StatePublished - Dec 9 1997
Externally publishedYes

Fingerprint

Peptide Fragments
Association reactions
Membranes
Peptides
Lipid bilayers
Lipid Bilayers
Immune Sera
Epitopes
Anti-Idiotypic Antibodies
Membrane Proteins
Kinetics
Antibodies

ASJC Scopus subject areas

  • Biochemistry

Cite this

Sheng, Z., Skach, W., Santarelli, V., & Deutsch, C. (1997). Evidence for interaction between transmembrane segments in assembly of Kv1.3. Biochemistry, 36(49), 15501-15513. https://doi.org/10.1021/bi971490j

Evidence for interaction between transmembrane segments in assembly of Kv1.3. / Sheng, ZuFang; Skach, William; Santarelli, Vincent; Deutsch, Carol.

In: Biochemistry, Vol. 36, No. 49, 09.12.1997, p. 15501-15513.

Research output: Contribution to journalArticle

Sheng, Z, Skach, W, Santarelli, V & Deutsch, C 1997, 'Evidence for interaction between transmembrane segments in assembly of Kv1.3', Biochemistry, vol. 36, no. 49, pp. 15501-15513. https://doi.org/10.1021/bi971490j
Sheng, ZuFang ; Skach, William ; Santarelli, Vincent ; Deutsch, Carol. / Evidence for interaction between transmembrane segments in assembly of Kv1.3. In: Biochemistry. 1997 ; Vol. 36, No. 49. pp. 15501-15513.
@article{aef354daafa341adbc5afdbdfdb25e14,
title = "Evidence for interaction between transmembrane segments in assembly of Kv1.3",
abstract = "Previously, we showed that the N-terminal recognition domain (T1) of Kv1.3 was not required for assembly of functional channels [Tu et al. (1996) J. Biol. Chem. 271, 18904-18911]. Moreover, specific Kv1.3 peptide fragments including regions of the central core are able to inhibit expression of current produced from a channel lacking the T1 domain, Kv1.3(T1-). To elucidate the mechanism whereby Kv1.3 peptide fragments suppress Kv1.3(T1-) current, we have studied the ability of peptide fragments containing the transmembrane segments S1, S1-S2, or S1-S2-S3 to physically associate with the Kv1.3(T1-) polypeptide subunit in vitro in microsomal membranes. Using c-myc (9E10) epitope-labeled peptide fragments and anti-myc antibody as well as antisera to the Kv1.3 C-terminus, we now demonstrate specific association of these peptide fragments with Kv1.3(T1-). Association of peptide fragments with Kv1.3(T1-) was correlated with integration of both proteins into the membrane. Furthermore, the relative strength and kinetics of this association directly correlated with the ability of fragments to suppress Kv1.3(T1-) current. The rate-limiting step in the sequential synthesis, integration, and formation of a complex was the association of integrated polypeptides within the plane of the lipid bilayer. These results strongly suggest that the physical association of transmembrane segments provides the basis for suppression of K+ channel function by K+ channel peptide fragments in vivo. Moreover, the S1-S2-S3 peptide fragment potently suppressed full-length Kv1.3, thus implicating a role for the S1-S2-S3 region of Kv1.3 in the assembly of the Kv1.3 channel. We refer to these putative association sites as IMA (intramembrane association) sites.",
author = "ZuFang Sheng and William Skach and Vincent Santarelli and Carol Deutsch",
year = "1997",
month = "12",
day = "9",
doi = "10.1021/bi971490j",
language = "English (US)",
volume = "36",
pages = "15501--15513",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "49",

}

TY - JOUR

T1 - Evidence for interaction between transmembrane segments in assembly of Kv1.3

AU - Sheng, ZuFang

AU - Skach, William

AU - Santarelli, Vincent

AU - Deutsch, Carol

PY - 1997/12/9

Y1 - 1997/12/9

N2 - Previously, we showed that the N-terminal recognition domain (T1) of Kv1.3 was not required for assembly of functional channels [Tu et al. (1996) J. Biol. Chem. 271, 18904-18911]. Moreover, specific Kv1.3 peptide fragments including regions of the central core are able to inhibit expression of current produced from a channel lacking the T1 domain, Kv1.3(T1-). To elucidate the mechanism whereby Kv1.3 peptide fragments suppress Kv1.3(T1-) current, we have studied the ability of peptide fragments containing the transmembrane segments S1, S1-S2, or S1-S2-S3 to physically associate with the Kv1.3(T1-) polypeptide subunit in vitro in microsomal membranes. Using c-myc (9E10) epitope-labeled peptide fragments and anti-myc antibody as well as antisera to the Kv1.3 C-terminus, we now demonstrate specific association of these peptide fragments with Kv1.3(T1-). Association of peptide fragments with Kv1.3(T1-) was correlated with integration of both proteins into the membrane. Furthermore, the relative strength and kinetics of this association directly correlated with the ability of fragments to suppress Kv1.3(T1-) current. The rate-limiting step in the sequential synthesis, integration, and formation of a complex was the association of integrated polypeptides within the plane of the lipid bilayer. These results strongly suggest that the physical association of transmembrane segments provides the basis for suppression of K+ channel function by K+ channel peptide fragments in vivo. Moreover, the S1-S2-S3 peptide fragment potently suppressed full-length Kv1.3, thus implicating a role for the S1-S2-S3 region of Kv1.3 in the assembly of the Kv1.3 channel. We refer to these putative association sites as IMA (intramembrane association) sites.

AB - Previously, we showed that the N-terminal recognition domain (T1) of Kv1.3 was not required for assembly of functional channels [Tu et al. (1996) J. Biol. Chem. 271, 18904-18911]. Moreover, specific Kv1.3 peptide fragments including regions of the central core are able to inhibit expression of current produced from a channel lacking the T1 domain, Kv1.3(T1-). To elucidate the mechanism whereby Kv1.3 peptide fragments suppress Kv1.3(T1-) current, we have studied the ability of peptide fragments containing the transmembrane segments S1, S1-S2, or S1-S2-S3 to physically associate with the Kv1.3(T1-) polypeptide subunit in vitro in microsomal membranes. Using c-myc (9E10) epitope-labeled peptide fragments and anti-myc antibody as well as antisera to the Kv1.3 C-terminus, we now demonstrate specific association of these peptide fragments with Kv1.3(T1-). Association of peptide fragments with Kv1.3(T1-) was correlated with integration of both proteins into the membrane. Furthermore, the relative strength and kinetics of this association directly correlated with the ability of fragments to suppress Kv1.3(T1-) current. The rate-limiting step in the sequential synthesis, integration, and formation of a complex was the association of integrated polypeptides within the plane of the lipid bilayer. These results strongly suggest that the physical association of transmembrane segments provides the basis for suppression of K+ channel function by K+ channel peptide fragments in vivo. Moreover, the S1-S2-S3 peptide fragment potently suppressed full-length Kv1.3, thus implicating a role for the S1-S2-S3 region of Kv1.3 in the assembly of the Kv1.3 channel. We refer to these putative association sites as IMA (intramembrane association) sites.

UR - http://www.scopus.com/inward/record.url?scp=0030662986&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030662986&partnerID=8YFLogxK

U2 - 10.1021/bi971490j

DO - 10.1021/bi971490j

M3 - Article

VL - 36

SP - 15501

EP - 15513

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 49

ER -