PMID- 26184075
OWN - NLM
STAT- MEDLINE
DCOM- 20160926
LR  - 20191210
IS  - 1756-2651 (Electronic)
IS  - 0021-924X (Print)
IS  - 0021-924X (Linking)
VI  - 159
IP  - 1
DP  - 2016 Jan
TI  - Interaction of a novel fluorescent GTP analogue with the small G-protein K-Ras.
PG  - 41-8
LID - 10.1093/jb/mvv071 [doi]
AB  - A novel fluorescent guanosine 5'-triphosphate (GTP) analogue, 
      2'(3')-O-6-(N-(7-nitrobenz-2-oxa-l,3-diazol-4-yl)amino) hexanoic-GTP (NBD-GTP), 
      was synthesized and utilized to monitor the effect of mutations in the functional 
      region of mouse K-Ras. The effects of the G12S, A59T and G12S/A59T mutations on 
      GTPase activity, nucleotide exchange rates were compared with normal Ras. 
      Mutation at A59T resulted in reduction of the GTPase activity by 0.6-fold and 
      enhancement of the nucleotide exchange rate by 2-fold compared with normal Ras. 
      On the other hand, mutation at G12S only slightly affected the nucleotide 
      exchange rate and did not affect the GTPase activity. We also used NBD-GTP to 
      study the effect of these mutations on the interaction between Ras and SOS1, a 
      guanine nucleotide exchange factor. The mutation at A59T abolished the 
      interaction with SOS1. The results suggest that the fluorescent GTP analogue, 
      NBD-GTP, is applicable to the kinetic studies for small G-proteins.
CI  - (c) The Authors 2015. Published by Oxford University Press on behalf of the 
      Japanese Biochemical Society. All rights reserved.
FAU - Iwata, Seigo
AU  - Iwata S
AD  - Division of Bioengineering, Graduate School of Engineering, Soka University, 
      Hachioji, Tokyo 192-8577, Japan;
FAU - Masuhara, Kaori
AU  - Masuhara K
AD  - Division of Bioengineering, Graduate School of Engineering, Soka University, 
      Hachioji, Tokyo 192-8577, Japan;
FAU - Umeki, Nobuhisa
AU  - Umeki N
AD  - Division of Bioengineering, Graduate School of Engineering, Soka University, 
      Hachioji, Tokyo 192-8577, Japan;
FAU - Sako, Yasushi
AU  - Sako Y
AD  - Division of Bioengineering, Graduate School of Engineering, Soka University, 
      Hachioji, Tokyo 192-8577, Japan;
FAU - Maruta, Shinsaku
AU  - Maruta S
AD  - Division of Bioengineering, Graduate School of Engineering, Soka University, 
      Hachioji, Tokyo 192-8577, Japan; Division of Bioengineering, Graduate School of 
      Engineering, Soka University, Hachioji, Tokyo 192-8577, Japan; maruta@soka.ac.jp.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20150715
PL  - England
TA  - J Biochem
JT  - Journal of biochemistry
JID - 0376600
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Guanine Nucleotide Exchange Factors)
RN  - 0 (SOS1 Protein)
RN  - 86-01-1 (Guanosine Triphosphate)
RN  - EC 3.6.5.2 (Hras protein, mouse)
RN  - EC 3.6.5.2 (Monomeric GTP-Binding Proteins)
RN  - EC 3.6.5.2 (Proto-Oncogene Proteins p21(ras))
RN  - EQF2794IRE (4-Chloro-7-nitrobenzofurazan)
SB  - IM
MH  - 4-Chloro-7-nitrobenzofurazan/*analogs & derivatives/chemical 
      synthesis/chemistry/metabolism
MH  - Animals
MH  - Fluorescent Dyes/chemical synthesis/chemistry/*metabolism
MH  - Guanine Nucleotide Exchange Factors/metabolism
MH  - Guanosine Triphosphate/*analogs & derivatives/chemical 
      synthesis/chemistry/*metabolism
MH  - Kinetics
MH  - Mice
MH  - Monomeric GTP-Binding Proteins/genetics/*metabolism
MH  - Mutation
MH  - Protein Binding
MH  - Proto-Oncogene Proteins p21(ras)/genetics/*metabolism
MH  - SOS1 Protein/metabolism
PMC - PMC4882638
OTO - NOTNLM
OT  - G-proteins
OT  - GEF
OT  - Ras
OT  - fluorescence
OT  - nucleotide analogue
EDAT- 2015/07/18 06:00
MHDA- 2016/09/27 06:00
PMCR- 2017/01/01
CRDT- 2015/07/18 06:00
PHST- 2015/04/27 00:00 [received]
PHST- 2015/06/09 00:00 [accepted]
PHST- 2015/07/18 06:00 [entrez]
PHST- 2015/07/18 06:00 [pubmed]
PHST- 2016/09/27 06:00 [medline]
PHST- 2017/01/01 00:00 [pmc-release]
AID - mvv071 [pii]
AID - 10.1093/jb/mvv071 [doi]
PST - ppublish
SO  - J Biochem. 2016 Jan;159(1):41-8. doi: 10.1093/jb/mvv071. Epub 2015 Jul 15.