Home LiteratureArticle Details
PMID: 2023911 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The first milliseconds of the pore formed by a fusogenic viral envelope protein during membrane fusion.

Spruce AE, Iwata A, Almers W

Abstract

Fibroblasts expressing the influenza virus hemagglutinin on their plasma membrane were patch clamped while they fused to erythrocytes. An increase in the fibroblast's membrane capacitance indicated the opening of the "fusion pore," the first aqueous connection between the fusing cells. We show here that the capacitance increase is preceded by a brief current transient, generated as the erythrocyte discharges its membrane potential through the nascent fusion pore. This signal allows one to calculate the pore conductance during the first milliseconds of its existence. The pore conductance jumps from 0 to approximately 150 pS and then grows more gradually over the subsequent tens of milliseconds until growth is arrested. The initial conductance is similar to that of a large ion channel and suggests that the pore is initially only 1-2 nm wide. Hence, we are probably observing events caused by only a small number of hemagglutinin molecules.

MeSH Terms
Animals Cell Membrane/physiology Electric Conductivity Erythrocytes/physiology Hemagglutinins, Viral/physiology Humans In Vitro Techniques Membrane Fusion Membrane Potentials Mice Recombinant Proteins Time Factors Viral Fusion Proteins/physiology
Chemicals
Hemagglutinins, Viral Recombinant Proteins Viral Fusion Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Spruce A E
Department of Physiology and Biophysics, University of Washington, Seattle 98195.
Iwata A
Almers W
References (19)
19 references, click to expand
  1. Three-dimensional model of purple membrane obtained by electron microscopy.
    Nature. 1975 Sep 4;257(5521):28-32 PMID: 1161000
  2. Fusion of influenza hemagglutinin-expressing fibroblasts with glycophorin-bearing liposomes: role of hemagglutinin surface density.
    Biochemistry. 1990 Oct 16;29(41):9697-707 PMID: 2271610
  3. Diameter of the cell-to-cell junctional membrane channels as probed with neutral molecules.
    Science. 1981 Jul 31;213(4507):551-3 PMID: 7244653
  4. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches.
    Pflugers Arch. 1981 Aug;391(2):85-100 PMID: 6270629
  5. Discrete changes of cell membrane capacitance observed under conditions of enhanced secretion in bovine adrenal chromaffin cells.
    Proc Natl Acad Sci U S A. 1982 Nov;79(21):6712-6 PMID: 6959149
  6. An efficient method for introducing macromolecules into living cells.
    J Cell Biol. 1985 Jul;101(1):19-27 PMID: 2989298
  7. Single-channel currents of an intercellular junction.
    Nature. 1985 Sep 26-Oct 2;317(6035):331-5 PMID: 2413362
  8. Fast events in single-channel currents activated by acetylcholine and its analogues at the frog muscle end-plate.
    J Physiol. 1985 Dec;369:501-57 PMID: 2419552
  9. Electron microscopy of the low pH structure of influenza virus haemagglutinin.
    EMBO J. 1986 Jan;5(1):41-9 PMID: 3956479
  10. Physiological modulation of gap junction permeability.
    J Exp Biol. 1986 Sep;124:993-114 PMID: 2428910
  11. The structure and function of the hemagglutinin membrane glycoprotein of influenza virus.
    Annu Rev Biochem. 1987;56:365-94 PMID: 3304138
  12. Currents through the fusion pore that forms during exocytosis of a secretory vesicle.
    Nature. 1987 Aug 27-Sep 2;328(6133):814-7 PMID: 2442614
  13. Cardiac gap junction channel activity in embryonic chick ventricle cells.
    Am J Physiol. 1988 Jan;254(1 Pt 2):H170-80 PMID: 3337253
  14. Protein-mediated membrane fusion.
    Annu Rev Biophys Biophys Chem. 1989;18:187-211 PMID: 2660823
  15. Initial stages of influenza hemagglutinin-induced cell fusion monitored simultaneously by two fluorescent events: cytoplasmic continuity and lipid mixing.
    J Cell Biol. 1989 Jul;109(1):113-22 PMID: 2745545
  16. Patch clamp studies of single cell-fusion events mediated by a viral fusion protein.
    Nature. 1989 Nov 30;342(6249):555-8 PMID: 2586627
  17. Viral and cellular membrane fusion proteins.
    Annu Rev Physiol. 1990;52:675-97 PMID: 2184772
  18. Properties of the fusion pore that forms during exocytosis of a mast cell secretory vesicle.
    Neuron. 1990 May;4(5):643-54 PMID: 2344404
  19. Chloride conductance of the amphiuma red cell membrane.
    J Membr Biol. 1978 Feb 6;39(1):27-48 PMID: 24748
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1991-05-01
Pages
3623-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC51504
Subset
IM
Grants
NIGMS NIH HHS · GM-39520 · United States
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com