PMID- 33109600
OWN - NLM
STAT- MEDLINE
DCOM- 20210323
LR  - 20240229
IS  - 1083-351X (Electronic)
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 295
IP  - 52
DP  - 2020 Dec 25
TI  - In crystallo screening for proline analog inhibitors of the proline cycle enzyme 
      PYCR1.
PG  - 18316-18327
LID - S0021-9258(17)50701-2 [pii]
LID - 10.1074/jbc.RA120.016106 [doi]
AB  - Pyrroline-5-carboxylate reductase 1 (PYCR1) catalyzes the biosynthetic 
      half-reaction of the proline cycle by reducing Delta(1)-pyrroline-5-carboxylate (P5C) 
      to proline through the oxidation of NAD(P)H. Many cancers alter their proline 
      metabolism by up-regulating the proline cycle and proline biosynthesis, and 
      knockdowns of PYCR1 lead to decreased cell proliferation. Thus, evidence is 
      growing for PYCR1 as a potential cancer therapy target. Inhibitors of cancer 
      targets are useful as chemical probes for studying cancer mechanisms and starting 
      compounds for drug discovery; however, there is a notable lack of validated 
      inhibitors for PYCR1. To fill this gap, we performed a small-scale focused screen 
      of proline analogs using X-ray crystallography. Five inhibitors of human PYCR1 
      were discovered: l-tetrahydro-2-furoic acid, cyclopentanecarboxylate, 
      l-thiazolidine-4-carboxylate, l-thiazolidine-2-carboxylate, and N-formyl 
      l-proline (NFLP). The most potent inhibitor was NFLP, which had a competitive 
      (with P5C) inhibition constant of 100 mum The structure of PYCR1 complexed with 
      NFLP shows that inhibitor binding is accompanied by conformational changes in the 
      active site, including the translation of an alpha-helix by 1 A. These changes are 
      unique to NFLP and enable additional hydrogen bonds with the enzyme. NFLP was 
      also shown to phenocopy the PYCR1 knockdown in MCF10A H-RAS(V12) breast cancer 
      cells by inhibiting de novo proline biosynthesis and impairing spheroidal growth. 
      In summary, we generated the first validated chemical probe of PYCR1 and 
      demonstrated proof-of-concept for screening proline analogs to discover 
      inhibitors of the proline cycle.
CI  - (c) 2020 Christensen et al.
FAU - Christensen, Emily M
AU  - Christensen EM
AD  - Department of Chemistry, University of Missouri, Columbia, Missouri, USA.
FAU - Bogner, Alexandra N
AU  - Bogner AN
AD  - Department of Biochemistry, University of Missouri, Columbia, Missouri, USA.
FAU - Vandekeere, Anke
AU  - Vandekeere A
AD  - Laboratory of Cellular Metabolism and Metabolic Regulation, VIB-KU Leuven Center 
      for Cancer Biology, VIB, Leuven, Belgium; Department of Oncology, Laboratory of 
      Cellular Metabolism and Metabolic Regulation, KU Leuven and Leuven Cancer 
      Institute (LKI), Leuven, Belgium.
FAU - Tam, Gabriela S
AU  - Tam GS
AD  - Department of Biochemistry, University of Missouri, Columbia, Missouri, USA.
FAU - Patel, Sagar M
AU  - Patel SM
AD  - Department of Biochemistry, Redox Biology Center, University of Nebraska, 
      Lincoln, Nebraska, USA.
FAU - Becker, Donald F
AU  - Becker DF
AD  - Department of Biochemistry, Redox Biology Center, University of Nebraska, 
      Lincoln, Nebraska, USA.
FAU - Fendt, Sarah-Maria
AU  - Fendt SM
AD  - Laboratory of Cellular Metabolism and Metabolic Regulation, VIB-KU Leuven Center 
      for Cancer Biology, VIB, Leuven, Belgium; Department of Oncology, Laboratory of 
      Cellular Metabolism and Metabolic Regulation, KU Leuven and Leuven Cancer 
      Institute (LKI), Leuven, Belgium.
FAU - Tanner, John J
AU  - Tanner JJ
AD  - Department of Biochemistry, University of Missouri, Columbia, Missouri, USA; 
      Department of Chemistry, University of Missouri, Columbia, Missouri, USA. 
      Electronic address: tannerjj@missouri.edu.
LA  - eng
SI  - PDB/5UAU
GR  - T32 GM008396/GM/NIGMS NIH HHS/United States
GR  - P30 GM124169/GM/NIGMS NIH HHS/United States
GR  - R01 GM132640/GM/NIGMS NIH HHS/United States
GR  - S10 OD021527/OD/NIH HHS/United States
GR  - P30 GM124165/GM/NIGMS NIH HHS/United States
GR  - R25 GM064120/GM/NIGMS 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 - 20201027
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Enzyme Inhibitors)
RN  - 9DLQ4CIU6V (Proline)
RN  - EC 1.5.1.- (Pyrroline Carboxylate Reductases)
SB  - IM
MH  - Breast Neoplasms/*metabolism/pathology
MH  - Catalytic Domain
MH  - Crystallography, X-Ray
MH  - Enzyme Inhibitors/*pharmacology
MH  - Female
MH  - Humans
MH  - Phenotype
MH  - Proline/*analogs & derivatives
MH  - Pyrroline Carboxylate Reductases/*antagonists & inhibitors/*metabolism
MH  - Tumor Cells, Cultured
MH  - delta-1-Pyrroline-5-Carboxylate Reductase
PMC - PMC7939384
OTO - NOTNLM
OT  - X-ray crystallography
OT  - breast cancer
OT  - enzyme inhibitor
OT  - enzyme kinetics
OT  - tumor metabolism
COIS- Conflict of interest-S.-M. F. has received funding from Bayer, Merck, and Black 
      Belt Therapeutics and has consulted for Fund+.
EDAT- 2020/10/29 06:00
MHDA- 2021/03/24 06:00
PMCR- 2021/01/13
CRDT- 2020/10/28 05:31
PHST- 2020/09/18 00:00 [received]
PHST- 2020/10/22 00:00 [revised]
PHST- 2020/10/29 06:00 [pubmed]
PHST- 2021/03/24 06:00 [medline]
PHST- 2020/10/28 05:31 [entrez]
PHST- 2021/01/13 00:00 [pmc-release]
AID - S0021-9258(17)50701-2 [pii]
AID - 10.1074/jbc.RA120.016106 [doi]
PST - ppublish
SO  - J Biol Chem. 2020 Dec 25;295(52):18316-18327. doi: 10.1074/jbc.RA120.016106. Epub 
      2020 Oct 27.