PMID- 26410361
OWN - NLM
STAT- MEDLINE
DCOM- 20161213
LR  - 20161230
IS  - 1638-6183 (Electronic)
IS  - 0300-9084 (Linking)
VI  - 123
DP  - 2016 Apr
TI  - Bifunctional activity of deoxyhypusine synthase/hydroxylase from Trichomonas 
      vaginalis.
PG  - 37-51
LID - S0300-9084(15)00302-8 [pii]
LID - 10.1016/j.biochi.2015.09.027 [doi]
AB  - The Trichomonas vaginalis genome analysis suggested the presence of a putative 
      deoxyhypusine synthase (TvDHS) that catalyzes the posttranslational modification 
      of eIF-5A. Herein, we expressed and purified the recombinant TvDHS (rTvDHS) 
      protein (43 kDa) and the recombinant TveIF-5A (rTveIF-5A) precursor protein 
      (46 kDa). A 41 kDa band of the native TvDHS was recognized by western blot 
      analysis in T. vaginalis total protein extract by a mouse polyclonal anti-rTvDHS 
      antibody. The enzymatic activity of rTvDHS was determined by in vitro rTveIF-5A 
      precursor modification. The modification reaction was performed by using 
      ((3)H)-spermidine, and the biochemical analysis showed that rTvDHS exhibited Km 
      value of 0.6 muM. The rTvDHS activity was inhibited by the spermidine analog, 
      N''-guanyl-1,7-diamino-heptane (GC7). Native gel electrophoresis analysis showed 
      two bands corresponding to an rTvDHS-rTveIF-5A complex and an intermediate form 
      of rTveIF-5A. The two forms were subsequently separated by ion exchange 
      chromatography to identify the hypusine residue by MS/MS analysis. Moreover, 
      mutations in TvDHS showed that the putative HE motif present in this enzyme is 
      involved in the hydroxylation of TveIF-5A. We observed that only 
      hypusine-containing TveIF-5A was bound to an RNA hairpin ERE structure from the 
      cox-2 gene, which contains the AAAUGUCACAC consensus sequence. Interestingly, 
      2DE-WB assays, using parasites that were grown in DAB-culture conditions and 
      transferred to exogenous putrescine, showed the new isoform of TveIF-5A. In 
      summary, our results indicate that T. vaginalis contains an active TvDHS capable 
      of modifying the precursor TveIF-5A protein, which subsequently exhibits RNA 
      binding activity.
CI  - Copyright (c) 2015. Published by Elsevier B.V.
FAU - Quintas-Granados, Laura Itzel
AU  - Quintas-Granados LI
AD  - Posgrado en Ciencias Genomicas, Universidad Autonoma de la Ciudad de Mexico 
      (UACM), San Lorenzo #290, Col. Del Valle, CP 03100, Mexico City, Mexico.
FAU - Carvajal Gamez, Bertha Isabel
AU  - Carvajal Gamez BI
AD  - Posgrado en Ciencias Genomicas, Universidad Autonoma de la Ciudad de Mexico 
      (UACM), San Lorenzo #290, Col. Del Valle, CP 03100, Mexico City, Mexico.
FAU - Villalpando, Jose Luis
AU  - Villalpando JL
AD  - Posgrado en Ciencias Genomicas, Universidad Autonoma de la Ciudad de Mexico 
      (UACM), San Lorenzo #290, Col. Del Valle, CP 03100, Mexico City, Mexico.
FAU - Ortega-Lopez, Jaime
AU  - Ortega-Lopez J
AD  - Departamento de Biotecnologia y Bioingenieria, Centro de Investigacion y de 
      Estudios Avanzados del IPN (CINVESTAV-IPN), Av. IPN 2508, Col. San Pedro 
      Zacatenco CP 07360, Mexico City, Mexico.
FAU - Arroyo, Rossana
AU  - Arroyo R
AD  - Departamento de Infectomica y Patogenesis Molecular, Centro de Investigacion y de 
      Estudios Avanzados del IPN (CINVESTAV-IPN), Av. IPN 2508, Col. San Pedro 
      Zacatenco CP 07360, Mexico City, Mexico.
FAU - Azuara-Liceaga, Elisa
AU  - Azuara-Liceaga E
AD  - Posgrado en Ciencias Genomicas, Universidad Autonoma de la Ciudad de Mexico 
      (UACM), San Lorenzo #290, Col. Del Valle, CP 03100, Mexico City, Mexico.
FAU - Alvarez-Sanchez, Maria Elizbeth
AU  - Alvarez-Sanchez ME
AD  - Posgrado en Ciencias Genomicas, Universidad Autonoma de la Ciudad de Mexico 
      (UACM), San Lorenzo #290, Col. Del Valle, CP 03100, Mexico City, Mexico. 
      Electronic address: elizbethalvarezsanchez@yahoo.com.mx.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20150926
PL  - France
TA  - Biochimie
JT  - Biochimie
JID - 1264604
RN  - EC 1.5.- (Oxidoreductases Acting on CH-NH Group Donors)
RN  - EC 1.5.1.- (deoxyhypusine synthase)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Chromatography, Liquid
MH  - Oxidoreductases Acting on CH-NH Group Donors/chemistry/*metabolism
MH  - Sequence Homology, Amino Acid
MH  - Spectrometry, Mass, Electrospray Ionization
MH  - Tandem Mass Spectrometry
MH  - Trichomonas vaginalis/*enzymology/genetics
OTO - NOTNLM
OT  - Bifunctional activity
OT  - Deoxyhypusine synthase
OT  - Eukaryotic translation initiation factor 5A
OT  - Eukaryotic-translation-initiation-factor-5A response element
OT  - Trichomonas vaginalis
EDAT- 2015/09/28 06:00
MHDA- 2016/12/15 06:00
CRDT- 2015/09/28 06:00
PHST- 2015/05/19 00:00 [received]
PHST- 2015/09/22 00:00 [accepted]
PHST- 2015/09/28 06:00 [entrez]
PHST- 2015/09/28 06:00 [pubmed]
PHST- 2016/12/15 06:00 [medline]
AID - S0300-9084(15)00302-8 [pii]
AID - 10.1016/j.biochi.2015.09.027 [doi]
PST - ppublish
SO  - Biochimie. 2016 Apr;123:37-51. doi: 10.1016/j.biochi.2015.09.027. Epub 2015 Sep 
      26.