PMID- 14970206
OWN - NLM
STAT- MEDLINE
DCOM- 20040610
LR  - 20220410
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 279
IP  - 17
DP  - 2004 Apr 23
TI  - Specific structural requirements for the inhibitory effect of thapsigargin on the 
      Ca2+ ATPase SERCA.
PG  - 17973-9
AB  - Mutational analysis of amino acid residues lining the thapsigargin (TG) binding 
      cavity at the interface of the membrane surface and cytosolic headpiece was 
      performed in the Ca(2+) ATPase (SERCA-1). Specific mutations such as F256V, 
      I765A, and Y837A reduce not only the apparent affinity of the ATPase for TG but 
      also the maximal inhibitory effect. The effect of mutations is dependent on the 
      type and size of the substitute side chain, indicating that hydrophobic 
      partitioning of TG and complementary molecular shapes are involved not only in 
      binding but also in the inhibitory mechanism. A major factor determining the 
      inhibitory effect of bound TG is its interference with conformational changes 
      that are required for the progress of the ATPase cycle. Most prominent and 
      specific is the TG interference with a wide displacement of the Phe-256 side 
      chain that is associated with the E2 to E1.2Ca(2+) transition. The specificity of 
      the TG inhibitory mechanism is emphasized by the finding that the F256V mutation 
      does not interfere at all with the effect of 2,5-di-(t-butyl)-hydroquinone, which 
      is another SERCA inhibitor bound by hydrophobic partitioning. The specificity of 
      the inhibitory mechanism is also emphasized by the observation that within the 
      concentration range producing total inhibition of wild-type SERCA-1, TG produces 
      a 4-fold stimulation of the P-glycoprotein (multidrug transporter) ATPase.
FAU - Xu, Cheng
AU  - Xu C
AD  - Department of Biochemistry, University of Maryland School of Medicine, Baltimore, 
      Maryland 21201-1503, USA.
FAU - Ma, Hailun
AU  - Ma H
FAU - Inesi, Giuseppe
AU  - Inesi G
FAU - Al-Shawi, Marwan K
AU  - Al-Shawi MK
FAU - Toyoshima, Chikashi
AU  - Toyoshima C
LA  - eng
GR  - R01-52502/PHS HHS/United States
GR  - R01HL69380/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
DEP - 20040217
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (ATP Binding Cassette Transporter, Subfamily B, Member 1)
RN  - 0 (DNA, Complementary)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Hydroquinones)
RN  - 0 (Lipid Bilayers)
RN  - 26XK13B61B (2,5-di-tert-butylhydroquinone)
RN  - 47E5O17Y3R (Phenylalanine)
RN  - 67526-95-8 (Thapsigargin)
RN  - EC 3.4.21.64 (Endopeptidase K)
RN  - EC 3.6.1.- (Adenosine Triphosphatases)
RN  - EC 3.6.3.8 (Sarcoplasmic Reticulum Calcium-Transporting ATPases)
RN  - EC 7.2.2.10 (Calcium-Transporting ATPases)
SB  - IM
MH  - ATP Binding Cassette Transporter, Subfamily B, Member 1/chemistry
MH  - Adenosine Triphosphatases/chemistry
MH  - Animals
MH  - Binding Sites
MH  - Blotting, Western
MH  - COS Cells
MH  - Calcium-Transporting ATPases/*chemistry/metabolism
MH  - Chickens
MH  - DNA Mutational Analysis
MH  - DNA, Complementary/metabolism
MH  - Dose-Response Relationship, Drug
MH  - Endopeptidase K/chemistry
MH  - Enzyme Inhibitors/pharmacology
MH  - Hydroquinones/pharmacology
MH  - Kinetics
MH  - Lipid Bilayers/metabolism
MH  - Models, Chemical
MH  - Models, Molecular
MH  - Muscle, Skeletal/metabolism
MH  - Mutation
MH  - Phenylalanine/chemistry
MH  - Protein Binding
MH  - Protein Conformation
MH  - Rabbits
MH  - Sarcoplasmic Reticulum/metabolism
MH  - Sarcoplasmic Reticulum Calcium-Transporting ATPases
MH  - Substrate Specificity
MH  - Thapsigargin/*pharmacology
MH  - Transfection
MH  - Transgenes
EDAT- 2004/02/19 05:00
MHDA- 2004/06/21 10:00
CRDT- 2004/02/19 05:00
PHST- 2004/02/19 05:00 [pubmed]
PHST- 2004/06/21 10:00 [medline]
PHST- 2004/02/19 05:00 [entrez]
AID - S0021-9258(19)75634-8 [pii]
AID - 10.1074/jbc.M313263200 [doi]
PST - ppublish
SO  - J Biol Chem. 2004 Apr 23;279(17):17973-9. doi: 10.1074/jbc.M313263200. Epub 2004 
      Feb 17.