Identification of the cardiac ryanodine receptor channel in membrane blebs of sarcoplasmic reticulum.

Abstract:

:Blebs of the sarcoplasmic reticulum (SR) membrane of heart muscle cells were generated after saponin perforation of the plasma membrane followed by complete hypercontraction of the cell. Although characteristic proteins of the plasma membrane, namely the beta1-adrenoreceptor and Galphai, were stained by monoclonal antibodies in the hypercontracted cells, these proteins could not be detected in the adjacent blebs. Monoclonal antibodies to the cardiac ryanodine receptor (RyR2), calsequestrin and SERCA2 bound at different amounts to surface components of the blebs and to components of the hypercontracted cells. From the immunofluorescence signals we conclude that the blebs are mainly constituted of corbular and junctional SR membrane, and only to a lesser extent of network SR membrane. Deconvolution microscopy revealed that the membrane location of RyR2, calsequestrin and SERCA2 in the bleb is comparable to native SR membrane. At the bleb membrane giga-ohm seals could be obtained and patches could be excised in a way that single-channel currents could be measured, although these are not completely identified.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Böhle T,Brandt MC,Henn N,Schmidt A,Bloch W,Beuckelmann DJ

doi

10.1016/s0014-5793(01)02862-9

keywords:

subject

Has Abstract

pub_date

2001-09-21 00:00:00

pages

419-25

issue

3

eissn

0014-5793

issn

1873-3468

pii

S0014-5793(01)02862-9

journal_volume

505

pub_type

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