PMID- 26297682
OWN - NLM
STAT- MEDLINE
DCOM- 20160421
LR  - 20191210
IS  - 1090-2449 (Electronic)
IS  - 0014-4894 (Linking)
VI  - 159
DP  - 2015 Dec
TI  - Presence of a thapsigargin-sensitive calcium pump in Trypanosoma evansi: 
      Immunological, physiological, molecular and structural evidences.
PG  - 107-17
LID - S0014-4894(15)30025-4 [pii]
LID - 10.1016/j.exppara.2015.08.017 [doi]
AB  - In higher eukaryotes, the sarco-endoplasmic reticulum (ER) Ca(2+)-ATPase (SERCA) 
      is characterized for its high sensitivity to low concentrations of thapsigargin 
      (TG), a very specific inhibitor. In contrast, SERCA-like enzymes with different 
      sensitivities to TG have been reported in trypanosomatids. Here, we characterized 
      a SERCA-like enzyme from Trypanosoma evansi and evaluated its interaction with 
      TG. Confocal fluorescence microscopy using BODIPY FL TG and specific anti-SERCA 
      antibodies localized the T. evansi SERCA-like enzyme in the ER and confirmed its 
      direct interaction with TG. Moreover, the use of either 1 muM TG or 25 muM 2',5'-di 
      (tert-butyl)-1,4-benzohydroquinone prevented the reuptake of Ca(2+) and 
      consequently produced a small increase in the parasite cytosolic calcium 
      concentration in a calcium-free medium, which was released from the ER pool. A 
      3035 bp-sequence coding for a protein with an estimated molecular mass of 
      110.2 kDa was cloned from T. evansi. The corresponding gene product contained all 
      the invariant residues and conserved motifs found in other P-type ATPases but 
      lacked the calmodulin binding site. Modeling of the three-dimensional structure 
      of the parasite enzyme revealed that the amino acid changes found in the TG-SERCA 
      binding pocket do not compromise the interaction between the enzyme and the 
      inhibitor. Therefore, we concluded that T. evansi possesses a SERCA-like protein 
      that is inhibited by TG.
CI  - Copyright (c) 2015 Elsevier Inc. All rights reserved.
FAU - Perez-Gordones, M C
AU  - Perez-Gordones MC
AD  - Instituto de Biologia Experimental (IBE), Universidad Central de Venezuela (UCV), 
      Caracas, Venezuela.
FAU - Serrano, M L
AU  - Serrano ML
AD  - Unidad de Quimica Medicinal, Facultad de Farmacia, Universidad Central de 
      Venezuela (UCV), Caracas, 1041A, Venezuela.
FAU - Rojas, H
AU  - Rojas H
AD  - Centro de Biofisica y Bioquimica (CBB), Instituto Venezolano de Investigaciones 
      Cientificas (IVIC), Caracas, Venezuela.
FAU - Martinez, J C
AU  - Martinez JC
AD  - Centro de Biociencias y Medicina Molecular, Instituto de Estudios Avanzados 
      (IDEA), Caracas, Venezuela.
FAU - Uzcanga, G
AU  - Uzcanga G
AD  - Centro de Biociencias y Medicina Molecular, Instituto de Estudios Avanzados 
      (IDEA), Caracas, Venezuela; Centro Internacional de Zoonosis (CIZ), Universidad 
      Central del Ecuador, Ecuador.
FAU - Mendoza, M
AU  - Mendoza M
AD  - Centro de Estudios Biomedicos y Veterinarios, Instituto de Estudios Cientificos y 
      Tecnologicos (IDECYT), Universidad Nacional Experimental Simon Bolivar, 
      Venezuela. Electronic address: mendozamarta17@gmail.com.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20150818
PL  - United States
TA  - Exp Parasitol
JT  - Experimental parasitology
JID - 0370713
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Ion Pumps)
RN  - 67526-95-8 (Thapsigargin)
RN  - EC 7.2.2.10 (Calcium-Transporting ATPases)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Blotting, Western
MH  - Calcium-Transporting ATPases/antagonists & inhibitors/*drug 
      effects/genetics/immunology
MH  - Endoplasmic Reticulum/enzymology
MH  - Enzyme Inhibitors/*pharmacology
MH  - Horse Diseases/parasitology
MH  - Horses
MH  - Ion Pumps/*drug effects/metabolism
MH  - Male
MH  - Microscopy, Confocal
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Sequence Alignment
MH  - Thapsigargin/*pharmacology
MH  - Trypanosoma/drug effects/*metabolism/physiology
MH  - Trypanosomiasis/parasitology/veterinary
OTO - NOTNLM
OT  - SERCA
OT  - Thapsigargin
OT  - Trypanosoma evansi
EDAT- 2015/08/25 06:00
MHDA- 2016/04/22 06:00
CRDT- 2015/08/23 06:00
PHST- 2015/03/09 00:00 [received]
PHST- 2015/07/28 00:00 [revised]
PHST- 2015/08/16 00:00 [accepted]
PHST- 2015/08/23 06:00 [entrez]
PHST- 2015/08/25 06:00 [pubmed]
PHST- 2016/04/22 06:00 [medline]
AID - S0014-4894(15)30025-4 [pii]
AID - 10.1016/j.exppara.2015.08.017 [doi]
PST - ppublish
SO  - Exp Parasitol. 2015 Dec;159:107-17. doi: 10.1016/j.exppara.2015.08.017. Epub 2015 
      Aug 18.