PMID- 30783001
OWN - NLM
STAT- MEDLINE
DCOM- 20200330
LR  - 20240213
IS  - 1098-6596 (Electronic)
IS  - 0066-4804 (Print)
IS  - 0066-4804 (Linking)
VI  - 63
IP  - 5
DP  - 2019 May
TI  - Enzymatic and Structural Characterization of the Naegleria fowleri Glucokinase.
LID - 10.1128/AAC.02410-18 [doi]
LID - e02410-18
AB  - Infection with the free-living amoeba Naegleria fowleri leads to life-threatening 
      primary amoebic meningoencephalitis. Efficacious treatment options for these 
      infections are limited, and the mortality rate is very high ( approximately 98%). Parasite 
      metabolism may provide suitable targets for therapeutic design. Like most other 
      organisms, glucose metabolism is critical for parasite viability, being required 
      for growth in culture. The first enzyme required for glucose metabolism is 
      typically a hexokinase (HK), which transfers a phosphate from ATP to glucose. The 
      products of this enzyme are required for both glycolysis and the pentose 
      phosphate pathway. However, the N. fowleri genome lacks an obvious HK homolog and 
      instead harbors a glucokinase (Glck). The N. fowleri Glck (NfGlck) shares limited 
      (25%) amino acid identity with the mammalian host enzyme (Homo sapiens Glck), 
      suggesting that parasite-specific inhibitors with anti-amoeba activity can be 
      generated. Following heterologous expression, NfGlck was found to have a limited 
      hexose substrate range, with the greatest activity observed with glucose. The 
      enzyme had apparent K(m) values of 42.5 +/- 7.3 muM and 141.6 +/- 9.9 muM for glucose 
      and ATP, respectively. The NfGlck structure was determined and refined to 2.2-A 
      resolution, revealing that the enzyme shares greatest structural similarity with 
      the Trypanosoma cruzi Glck. These similarities include binding modes and binding 
      environments for substrates. To identify inhibitors of NfGlck, we screened a 
      small collection of inhibitors of glucose-phosphorylating enzymes and identified 
      several small molecules with 50% inhibitory concentration values of <1 muM that 
      may prove useful as hit chemotypes for further leads and therapeutic development 
      against N. fowleri.
CI  - Copyright (c) 2019 American Society for Microbiology.
FAU - Milanes, Jillian E
AU  - Milanes JE
AD  - Eukaryotic Pathogens Innovation Center, Department of Genetics and Biochemistry, 
      Clemson University, Clemson, South Carolina, USA.
FAU - Suryadi, Jimmy
AU  - Suryadi J
AD  - Eukaryotic Pathogens Innovation Center, Department of Genetics and Biochemistry, 
      Clemson University, Clemson, South Carolina, USA.
FAU - Abendroth, Jan
AU  - Abendroth J
AD  - UCB Seattle/Seattle Structural Genomics Center for Infection Disease, Bainbridge 
      Island, Washington, USA.
FAU - Van Voorhis, Wesley C
AU  - Van Voorhis WC
AD  - Center for Emerging and Re-emerging Infectious Diseases Division of Allergy and 
      Infectious Diseases, Department of Medicine, University of Washington, Seattle, 
      Washington, USA.
FAU - Barrett, Kayleigh F
AU  - Barrett KF
AD  - Center for Emerging and Re-emerging Infectious Diseases Division of Allergy and 
      Infectious Diseases, Department of Medicine, University of Washington, Seattle, 
      Washington, USA.
FAU - Dranow, David M
AU  - Dranow DM
AD  - UCB Seattle/Seattle Structural Genomics Center for Infection Disease, Bainbridge 
      Island, Washington, USA.
FAU - Phan, Isabelle Q
AU  - Phan IQ
AD  - Seattle Structural Genomics Center for Infectious Disease, Center for Global 
      Infection Disease Research Seattle Children's Research Institute, Seattle, 
      Washington, USA.
FAU - Patrick, Stephen L
AU  - Patrick SL
AD  - Eukaryotic Pathogens Innovation Center, Department of Genetics and Biochemistry, 
      Clemson University, Clemson, South Carolina, USA.
FAU - Rozema, Soren D
AU  - Rozema SD
AD  - School of Pharmacy Pharmaceutical Sciences Division, University of 
      Wisconsin-Madison, Madison, Wisconsin, USA.
FAU - Khalifa, Muhammad M
AU  - Khalifa MM
AD  - School of Pharmacy Pharmaceutical Sciences Division, University of 
      Wisconsin-Madison, Madison, Wisconsin, USA.
FAU - Golden, Jennifer E
AU  - Golden JE
AUID- ORCID: 0000-0002-6813-3710
AD  - School of Pharmacy Pharmaceutical Sciences Division, University of 
      Wisconsin-Madison, Madison, Wisconsin, USA.
FAU - Morris, James C
AU  - Morris JC
AD  - Eukaryotic Pathogens Innovation Center, Department of Genetics and Biochemistry, 
      Clemson University, Clemson, South Carolina, USA jmorri2@clemson.edu.
LA  - eng
GR  - HHSN272201700059C/AI/NIAID NIH HHS/United States
GR  - P20 GM109094/GM/NIGMS NIH HHS/United States
GR  - HHSN271201700059C/DA/NIDA NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20190425
PL  - United States
TA  - Antimicrob Agents Chemother
JT  - Antimicrobial agents and chemotherapy
JID - 0315061
RN  - 0 (Protozoan Proteins)
RN  - 8L70Q75FXE (Adenosine Triphosphate)
RN  - EC 2.7.1.2 (Glucokinase)
RN  - IY9XDZ35W2 (Glucose)
SB  - IM
MH  - Adenosine Triphosphate/metabolism
MH  - Animals
MH  - Glucokinase/*chemistry/*metabolism
MH  - Glucose/metabolism
MH  - Humans
MH  - Naegleria fowleri/*enzymology
MH  - Protozoan Proteins/*chemistry/*metabolism
MH  - Trypanosoma cruzi/enzymology
PMC - PMC6496100
OTO - NOTNLM
OT  - Naegleria fowleri
OT  - glucokinase
EDAT- 2019/02/21 06:00
MHDA- 2020/03/31 06:00
PMCR- 2019/10/25
CRDT- 2019/02/21 06:00
PHST- 2018/11/15 00:00 [received]
PHST- 2019/02/08 00:00 [accepted]
PHST- 2019/02/21 06:00 [pubmed]
PHST- 2020/03/31 06:00 [medline]
PHST- 2019/02/21 06:00 [entrez]
PHST- 2019/10/25 00:00 [pmc-release]
AID - AAC.02410-18 [pii]
AID - 02410-18 [pii]
AID - 10.1128/AAC.02410-18 [doi]
PST - epublish
SO  - Antimicrob Agents Chemother. 2019 Apr 25;63(5):e02410-18. doi: 
      10.1128/AAC.02410-18. Print 2019 May.