PMID- 22040412
OWN - NLM
STAT- MEDLINE
DCOM- 20120501
LR  - 20181109
IS  - 2542-5641 (Electronic)
IS  - 0366-6999 (Linking)
VI  - 124
IP  - 17
DP  - 2011 Sep
TI  - Virtual mutagenesis of isocitrate dehydrogenase 1 involved in glioblastoma 
      multiforme.
PG  - 2611-5
AB  - BACKGROUND: Site A132Arg mutations potentially impair the affinity of isocitrate 
      dehydrogenase 1 (IDH1) for its substrate isocitrate (ICT), consequently reducing 
      the production of alpha-ketoglutarate and leading to tumor growth through the 
      induction of the hypoxia-inducible factor-1 (HIF-1) pathway. However, given that 
      the roles of other active sites in IDH1 substrate binding remain unclear, we 
      aimed to investigate IDH1 mutation pattern and its influence on enzyme function. 
      METHODS: Fifteen IDH1 catalytic active site candidates were selected for in 
      silico mutagenesis and protein homology modeling. Binding free energy of the 
      IDH1/ICT complexes with single-site mutations was compared with that of the wild 
      type. The affinity of 10 IDH1 catalytic active sites for the ICT substrate was 
      further calculated. RESULTS: The IDH1 active site included seven residues from 
      chain A (A77Thr, A94Ser, A100Arg, A132Arg, A109Arg, A275Asp, and A279Asp) and 
      three residues from chain B (B214Thr, B212Lys, and B252Asp) that constituted the 
      substrate ICT-binding site. These residues were located within 0.5 nm of ICT, 
      indicating a potential interaction with the substrate. IDH1 changes of binding 
      free energy (DeltaE) suggested that the A132Arg residue from chain A contributes 
      three hydrogen bonds to the ICT alpha-carboxyl and beta-carboxyl groups, while the other 
      nine residues involved in ICT binding form only one or two hydrogen bonds. Amino 
      acid substitutes at A132Arg, A109Arg, and B212Lys sites, had the greatest effect 
      on enzyme affinity for its substrate. CONCLUSIONS: Mutations at sites A132Arg, 
      A109Arg, and B212Lys reduced IDH1 affinity for ICT, indicating these active sites 
      may play a central role in substrate binding. Mutations at sites A77Thr, A94Ser, 
      and A275Asp increased the affinity of IDH1 for ICT, which may enhance IDN1 
      catalytic activity. Mutant IDH1 proteins with higher catalytic activity than the 
      wild-type IDH1 could potentially be used as a novel gene therapy for glioblastoma 
      multiforme.
FAU - Wang, Ming-dong
AU  - Wang MD
AD  - Department of Neurosurgery, Affiliated Hospital of Hebei University, Baoding, 
      Hebei 071000, China.
FAU - Shi, Yan-fang
AU  - Shi YF
FAU - Wang, Hong
AU  - Wang H
FAU - Wang, Jia-liang
AU  - Wang JL
FAU - Ma, Wen-bin
AU  - Ma WB
FAU - Wang, Ren-zhi
AU  - Wang RZ
LA  - eng
PT  - Journal Article
PL  - China
TA  - Chin Med J (Engl)
JT  - Chinese medical journal
JID - 7513795
RN  - 0 (Isocitrates)
RN  - 9RW6G5D4MQ (isocitric acid)
RN  - EC 1.1.1.41 (Isocitrate Dehydrogenase)
SB  - IM
MH  - Catalytic Domain/genetics
MH  - Glioblastoma/*genetics
MH  - Humans
MH  - Isocitrate Dehydrogenase/*genetics/*metabolism
MH  - Isocitrates/metabolism
MH  - Mutagenesis
MH  - Mutation
MH  - Protein Binding
MH  - Structure-Activity Relationship
EDAT- 2011/11/02 06:00
MHDA- 2012/05/02 06:00
CRDT- 2011/11/02 06:00
PHST- 2011/11/02 06:00 [entrez]
PHST- 2011/11/02 06:00 [pubmed]
PHST- 2012/05/02 06:00 [medline]
PST - ppublish
SO  - Chin Med J (Engl). 2011 Sep;124(17):2611-5.