Locating the thapsigargin-binding site on Ca(2+)-ATPase by cryoelectron microscopy

Journal of Molecular Biology
H S YoungDavid L Stokes

Abstract

Thapsigargin (TG) is a potent inhibitor of Ca(2+)-ATPase from sarcoplasmic and endoplasmic reticula. Previous enzymatic studies have concluded that Ca(2+)-ATPase is locked in a dead-end complex upon binding TG with an affinity of <1 nM and that this complex closely resembles the E(2) enzymatic state. We have studied the structural effects of TG binding by cryoelectron microscopy of tubular crystals, which have previously been shown to comprise Ca(2+)-ATPase molecules in the E(2) conformation. In particular, we have compared 3D reconstructions of Ca(2+)-ATPase in the absence and presence of either TG or its dansylated derivative. The overall molecular shape of Ca(2+)-ATPase in the reconstructions is very similar, demonstrating that the TG/Ca(2+)-ATPase complex does indeed physically resemble the E(2) conformation, in contrast to massive domain movements that appear to be induced by Ca(2+) binding. Difference maps reveal a consistent difference on the lumenal side of the membrane, which we conclude corresponds to the thapsigargin-binding site. Modeling the atomic structure for Ca(2+)-ATPase into our density maps reveals that this binding site is composed of the loops between transmembrane segments M3/M4 and M7/M8. Indirect effect...Continue Reading

References

Jul 26, 1976·Biochemical and Biophysical Research Communications·Y Dupont
Jun 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·S M Gruner
Mar 1, 1974·Proceedings of the National Academy of Sciences of the United States of America·G B WarrenJ C Metcalfe
Dec 1, 1984·Journal of Bioenergetics and Biomembranes·A Maurer, S Fleischer
Jul 1, 1994·Biophysical Journal·J VossD D Thomas
May 6, 1996·Biochimica Et Biophysica Acta·J V MøllerM le Maire
Mar 1, 1997·Biophysical Journal·K YonekuraC Toyoshima
Mar 7, 1998·Journal of Molecular Evolution·K B Axelsen, M G Palmgren

❮ Previous
Next ❯

Citations

May 4, 2011·World Journal of Biological Chemistry·Jyoti PandeAshok K Grover
Feb 25, 2003·Annual Review of Biophysics and Biomolecular Structure·David L Stokes, N Michael Green
May 17, 2018·The Journal of Biological Chemistry·Olga N RaguimovaSeth L Robia
May 11, 2007·The Journal of Biological Chemistry·Morad OueslatiRalf Schülein
Mar 1, 2005·The Journal of Biological Chemistry·David L StokesJean-Jacques Lacapère
Jan 30, 2007·The Journal of Biological Chemistry·Karine MoncoqHoward S Young
Feb 7, 2002·Journal of Molecular Biology·Chen XuDavid L Stokes
Jan 15, 2005·Brain Research·Alexandra CorralesEsperanza Recio-Pinto

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.