| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
J. Biol. Chem., Vol. 278, Issue 44, 43781-43786, October 31, 2003
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
From the
Department Pharmazie-Pharmakologie für Naturwissenschaften, Ludwig-Maximilians Universität München, Butenandtstrasse 7, 81377 München, Germany and the Institut für Pharmakologie und Toxikologie der Technischen, Universität München, Biedersteinerstrasse 29, 80802 München, Germany
The coassembly of homologous subunits to heteromeric complexes serves as an important mechanism in generating ion channel diversity. Here, we have studied heteromerization in the hyperpolarization-activated cyclic nucleotide-gated (HCN) channel family. Using a combination of fluorescence confocal microscopy, coimmunoprecipitation, and electrophysiology we found that upon coexpression in HEK293 cells almost all dimeric combinations of HCN channel subunits give rise to the formation of stable channel complexes in the plasma membrane. We also identified HCN1/HCN2 heteromers in mouse brain indicating that heteromeric channels exist in vivo. Surprisingly, HCN2 and HCN3 did not coassemble to heteromeric channels. This finding indicates that heteromerization requires specific structural determinants that are not present in all HCN channel combinations. Using N-glycosidase F we show that native as well as recombinant HCN channels are glycosylated resulting in a 10–20-kDa shift in the molecular weight. Tunicamycin, an inhibitor of N-linked glycosylation, blocked surface membrane expression of HCN2. Similarly, a mutant HCN2 channel in which the putative N-glycosylation site in the loop between S5 and the pore helix was replaced by glutamine (HCN2N380Q) was not inserted into the plasma membrane and did not yield detectable whole-cell currents. These results indicate that N-linked glycosylation is required for cell surface trafficking of HCN channels. Cotransfection of HCN2N380Q with HCN4, but not with HCN3, rescued cell surface expression of HCN2N380Q. Immunoprecipitation revealed that this rescue was due to the formation of a HCN2N380Q/HCN4 heteromeric channel. Taken together our results indicate that subunit heteromerization and glycosylation are important determinants of the formation of native HCN channels.
Received for publication, June 30, 2003 , and in revised form, August 6, 2003.
* This work was supported by grants from the Deutsche Forschungsgemeinschaft. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. Pharmazie-Pharmakologie für Naturwissenschaften, Ludwig-Maximilians Universität München, Butenandtstr. 7, 81377 München, Germany. Tel.: 49-89-2180-77327; Fax: 49-89-2180-77326; E-mail: mbiel{at}cup.uni-muenchen.de.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  C.-H. Li, Q. Zhang, B. Teng, S. J. Mustafa, J.-Y. Huang, and H.-G. Yu Src tyrosine kinase alters gating of hyperpolarization-activated HCN4 pacemaker channel through Tyr531 Am J Physiol Cell Physiol, January 1, 2008; 294(1): C355 - C362. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S.-W. Ying, F. Jia, S. Y. Abbas, F. Hofmann, A. Ludwig, and P. A. Goldstein Dendritic HCN2 Channels Constrain Glutamate-Driven Excitability in Reticular Thalamic Neurons J. Neurosci., August 8, 2007; 27(32): 8719 - 8732. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. M. Whitaker, D. Angoli, H. Nazzari, R. Shigemoto, and E. A. Accili HCN2 and HCN4 Isoforms Self-assemble and Co-assemble with Equal Preference to Form Functional Pacemaker Channels J. Biol. Chem., August 3, 2007; 282(31): 22900 - 22909. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. A. Jackson, C. R. Marshall, and E. A. Accili Evolution and structural diversification of hyperpolarization-activated cyclic nucleotide-gated channel genes Physiol Genomics, May 11, 2007; 29(3): 231 - 245. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. A. Bender, T. Kirschstein, O. Kretz, A. L. Brewster, C. Richichi, C. Ruschenschmidt, R. Shigemoto, H. Beck, M. Frotscher, and T. Z. Baram Localization of HCN1 Channels to Presynaptic Compartments: Novel Plasticity That May Contribute to Hippocampal Maturation J. Neurosci., April 25, 2007; 27(17): 4697 - 4706. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. L. Brewster, Y. Chen, R. A. Bender, A. Yeh, R. Shigemoto, and T. Z. Baram Quantitative Analysis and Subcellular Distribution of mRNA and Protein Expression of the Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels throughout Development in Rat Hippocampus Cereb Cortex, March 1, 2007; 17(3): 702 - 712. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Mochizuki, T. Kagawa, A. Numari, M. J. Harris, J. Itoh, N. Watanabe, T. Mine, and I. M. Arias Two N-linked glycans are required to maintain the transport activity of the bile salt export pump (ABCB11) in MDCK II cells Am J Physiol Gastrointest Liver Physiol, March 1, 2007; 292(3): G818 - G828. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Erler, D. M. M. Al-Ansary, U. Wissenbach, T. F. J. Wagner, V. Flockerzi, and B. A. Niemeyer Trafficking and Assembly of the Cold-sensitive TRPM8 Channel J. Biol. Chem., December 15, 2006; 281(50): 38396 - 38404. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Michalakis, J. Reisert, H. Geiger, C. Wetzel, X. Zong, J. Bradley, M. Spehr, S. Huttl, A. Gerstner, A. Pfeifer, et al. Loss of CNGB1 Protein Leads to Olfactory Dysfunction and Subciliary Cyclic Nucleotide-gated Channel Trapping J. Biol. Chem., November 17, 2006; 281(46): 35156 - 35166. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. E. Leao, R. N. Leao, H. Sun, R. E. W. Fyffe, and B. Walmsley Hyperpolarization-activated currents are differentially expressed in mice brainstem auditory nuclei J. Physiol., November 1, 2006; 576(3): 849 - 864. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Xu, Y. Fu, W. Tian, and D. M. Cohen Glycosylation of the osmoresponsive transient receptor potential channel TRPV4 on Asn-651 influences membrane trafficking Am J Physiol Renal Physiol, May 1, 2006; 290(5): F1103 - F1109. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Bucchi, A. Tognati, R. Milanesi, M. Baruscotti, and D. DiFrancesco Properties of ivabradine-induced block of HCN1 and HCN4 pacemaker channels J. Physiol., April 15, 2006; 572(2): 335 - 346. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X. Chen, S. Shu, and D. A. Bayliss Suppression of Ih Contributes to Propofol-Induced Inhibition of Mouse Cortical Pyramidal Neurons J Neurophysiol, December 1, 2005; 94(6): 3872 - 3883. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Hofmann, M. Biel, and U. B. Kaupp International Union of Pharmacology. LI. Nomenclature and Structure-Function Relationships of Cyclic Nucleotide-Regulated Channels Pharmacol. Rev., December 1, 2005; 57(4): 455 - 462. [Full Text] [PDF]
|
|
|
|
|
|
T. Budde, L. Caputi, T. Kanyshkova, R. Staak, C. Abrahamczik, T. Munsch, and H.-C. Pape Impaired Regulation of Thalamic Pacemaker Channels through an Imbalance of Subunit Expression in Absence Epilepsy J. Neurosci., October 26, 2005; 25(43): 9871 - 9882. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X. Zong, C. Eckert, H. Yuan, C. Wahl-Schott, H. Abicht, L. Fang, R. Li, P. Mistrik, A. Gerstner, B. Much, et al. A Novel Mechanism of Modulation of Hyperpolarization-activated Cyclic Nucleotide-gated Channels by Src Kinase J. Biol. Chem., October 7, 2005; 280(40): 34224 - 34232. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Mistrik, R. Mader, S. Michalakis, M. Weidinger, A. Pfeifer, and M. Biel The Murine HCN3 Gene Encodes a Hyperpolarization-activated Cation Channel with Slow Kinetics and Unique Response to Cyclic Nucleotides J. Biol. Chem., July 22, 2005; 280(29): 27056 - 27061. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Akhavan, R. Atanasiu, T. Noguchi, W. Han, N. Holder, and A. Shrier Identification of the cyclic-nucleotide-binding domain as a conserved determinant of ion-channel cell-surface localization J. Cell Sci., July 1, 2005; 118(13): 2803 - 2812. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X. Chen, J. E. Sirois, Q. Lei, E. M. Talley, C. Lynch III, and D. A. Bayliss HCN Subunit-Specific and cAMP-Modulated Effects of Anesthetics on Neuronal Pacemaker Currents J. Neurosci., June 15, 2005; 25(24): 5803 - 5814. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Wahl-Schott, L. Baumann, X. Zong, and M. Biel An Arginine Residue in the Pore Region Is a Key Determinant of Chloride Dependence in Cardiac Pacemaker Channels J. Biol. Chem., April 8, 2005; 280(14): 13694 - 13700. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Michalakis, H. Geiger, S. Haverkamp, F. Hofmann, A. Gerstner, and M. Biel Impaired Opsin Targeting and Cone Photoreceptor Migration in the Retina of Mice Lacking the Cyclic Nucleotide-Gated Channel CNGA3 Invest. Ophthalmol. Vis. Sci., April 1, 2005; 46(4): 1516 - 1524. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Huttl, S. Michalakis, M. Seeliger, D.-G. Luo, N. Acar, H. Geiger, K. Hudl, R. Mader, S. Haverkamp, M. Moser, et al. Impaired Channel Targeting and Retinal Degeneration in Mice Lacking the Cyclic Nucleotide-Gated Channel Subunit CNGB1 J. Neurosci., January 5, 2005; 25(1): 130 - 138. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Qu, Y. Kryukova, I. A. Potapova, S. V. Doronin, M. Larsen, G. Krishnamurthy, I. S. Cohen, and R. B. Robinson MiRP1 Modulates HCN2 Channel Expression and Gating in Cardiac Myocytes J. Biol. Chem., October 15, 2004; 279(42): 43497 - 43502. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. P. Faillace, R. O. Bernabeu, and J. I. Korenbrot Cellular Processing of Cone Photoreceptor Cyclic GMP-gated Ion Channels: A ROLE FOR THE S4 STRUCTURAL MOTIF J. Biol. Chem., May 21, 2004; 279(21): 22643 - 22653. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
