Role of the sequence surrounding predicted transmembrane helix M4 in membrane association and function of the Ca(2+) release channel of skeletal muscle sarcoplasmic reticulum (ryanodine receptor isoform 1)

The Journal of Biological Chemistry
Guo Guang DuD H MacLennan

Abstract

The role of the sequence surrounding M4 in ryanodine receptors (RyR) in membrane association and function was investigated. This sequence contains a basic, 19-amino acid M3/M4 loop, a hydrophobic 44-49 amino acid sequence designated M4 (or M4a/M4b), and a hydrophilic M4/M5 loop. Enhanced green fluorescent protein (EGFP) was inserted into RyR1 and truncated just after the basic sequence, just after M4, within the M4/M5 loop, just before M5 and just after M5. The A52 epitope was inserted into RyR2 and truncated just after M4a. Analysis of these constructs ruled out a M3/M4 transmembrane hairpin and narrowed the region of membrane association to M4a/M4b. EGFP inserted between M4a and M4b in full-length RyR2 was altered conformationally, losing fluorescence and gaining trypsin sensitivity. Although it was accessible to an antibody from the cytosolic side, tryptic fragments were membrane-bound. The expressed protein containing EGFP retained caffeine-induced Ca(2+) release channel function. These results suggest that M4a/M4b either forms a transmembrane hairpin or associates in an unorthodox fashion with the cytosolic leaflet of the membrane, possibly involving the basic M3/M4 loop. The expression of a mutant RyR1, Delta4274-4535, de...Continue Reading

References

Jan 30, 1986·Biochimica Et Biophysica Acta·J K Mohana Rao, P Argos
May 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·K Maruyama, D H MacLennan
Aug 1, 1987·Molecular and Cellular Biology·C Chen, H Okayama
Jan 1, 1986·Methods in Enzymology·P Argos, J K Rao
Jan 1, 1973·Journal of Supramolecular Structure·T L Steck, J Yu
Aug 31, 1981·Biochemical and Biophysical Research Communications·S H Bokman, W W Ward
Mar 25, 1994·Journal of Molecular Biology·B Persson, P Argos
Mar 1, 1993·Trends in Pharmacological Sciences·V Sorrentino, P Volpe
Oct 17, 1998·The Journal of Biological Chemistry·K OtaK Mihara
Oct 3, 1999·The Journal of Biological Chemistry·D L GalvanG A Mignery
Mar 29, 2000·The Journal of General Physiology·G Avila, R T Dirksen
Mar 30, 2001·The Journal of Biological Chemistry·G AvilaR T Dirksen
Dec 18, 2002·Proceedings of the National Academy of Sciences of the United States of America·Guo Guang DuDavid H MacLennan
May 2, 2003·Nature·Youxing JiangRoderick MacKinnon
May 2, 2003·Nature·Youxing JiangRoderick MacKinnon

❮ Previous
Next ❯

Citations

Feb 7, 2008·European Biophysics Journal : EBJ·Irina BaranVirgil Baran
Jan 16, 2013·The Journal of Biological Chemistry·Srinivas RamachandranGerhard Meissner
Apr 25, 2012·Clinical and Experimental Pharmacology & Physiology·Angela F DulhuntyNicole A Beard
Jul 26, 2011·Science China. Life Sciences·Lynn Kimlicka, Filip Van Petegem
Oct 13, 2005·Cell Calcium·Ulrich Ebbinghaus-KintscherMasanori Tohnishi
Aug 24, 2005·Cell Calcium·Susan L Hamilton
Aug 9, 2005·Structure·Steven J LudtkeWah Chiu
May 29, 2007·The Journal of Biological Chemistry·Gargi MeurColin W Taylor
Sep 6, 2008·Circulation Journal : Official Journal of the Japanese Circulation Society·Masafumi YanoMasunori Matsuzaki
Jun 6, 2020·The Journal of Biological Chemistry·Moran ElbazFrancesco Zorzato

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.