Structural and functional insight into serine hydroxymethyltransferase from Helicobacter pylori

Autor: Hannu Myllykallio, Andreea Sodolescu, Ursula Liebl, Stéphane Skouloubris, Laurent Terradot, Latifa Bouzhir-Sima, Cyril Dian, Roxane Lestini
Přispěvatelé: Institute for Integrative Biology of the Cell, Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Institut de biologie et chimie des protéines [Lyon] (IBCP), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'optique et biosciences (LOB), École polytechnique (X)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut de Biologie Intégrative de la Cellule (I2BC), Institut des sciences du végétal (ISV), Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Optique et Biosciences (LOB), Université Paris-Sud - Paris 11 (UP11)
Jazyk: angličtina
Rok vydání: 2018
Předmět:
0301 basic medicine
[SDV]Life Sciences [q-bio]
Artificial Gene Amplification and Extension
Pathology and Laboratory Medicine
Thymidylate synthase
Biochemistry
Polymerase Chain Reaction
Serine
chemistry.chemical_compound
Helicobacter
Dihydrofolate reductase
Medicine and Health Sciences
Transferase
Amino Acids
Enzyme Chemistry
ComputingMilieux_MISCELLANEOUS
Glycine Hydroxymethyltransferase
Multidisciplinary
Crystallography
biology
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Structural Biology [q-bio.BM]
Organic Compounds
Physics
Biochemical Cofactors
Monomers
Condensed Matter Physics
Enzyme structure
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Biomolecules [q-bio.BM]
Bacterial Pathogens
Chemistry
Medical Microbiology
Physical Sciences
Crystal Structure
Medicine
Pathogens
Research Article
Science
Glycine
Research and Analysis Methods
Microbiology
Catalysis
03 medical and health sciences
Folic Acid
Biosynthesis
Bacterial Proteins
Protein Domains
Hydroxyl Amino Acids
Escherichia coli
Solid State Physics
[SDV.BBM]Life Sciences [q-bio]/Biochemistry
Molecular Biology
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Biochemistry [q-bio.BM]
Molecular Biology Techniques
Microbial Pathogens
Molecular Biology
Bacteria
Helicobacter pylori
Organic Chemistry
Genetic Complementation Test
Organisms
Chemical Compounds
Biology and Life Sciences
Proteins
[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Molecular biology
Polymer Chemistry
030104 developmental biology
chemistry
Aliphatic Amino Acids
Serine hydroxymethyltransferase
Enzyme Structure
biology.protein
Enzymology
Zdroj: PLoS ONE, Vol 13, Iss 12, p e0208850 (2018)
PLoS ONE
PLoS ONE, Public Library of Science, 2018, 13 (12), pp.e0208850. ⟨10.1371/journal.pone.0208850⟩
PLoS ONE, 2018, 13 (12), pp.e0208850. ⟨10.1371/journal.pone.0208850⟩
'PloS One ', vol: 13, pages: e0208850-1-e0208850-23 (2018)
ISSN: 1932-6203
DOI: 10.1371/journal.pone.0208850⟩
Popis: Serine hydroxymethyltransferase (SHMT), encoded by the glyA gene, is a ubiquitous pyridoxal 5’-phosphate (PLP)-dependent enzyme that catalyzes the formation of glycine from serine. The thereby generated 5,10-methylene tetrahydrofolate (MTHF) is a major source of cellular one-carbon units and a key intermediate in thymidylate biosynthesis. While in virtually all eukaryotic and many bacterial systems thymidylate synthase ThyA, SHMT and dihydrofolate reductase (DHFR) are part of the thymidylate/folate cycle, the situation is different in organisms using flavin-dependent thymidylate synthase ThyX. Here the distinct catalytic reaction directly produces tetrahydrofolate (THF) and consequently in most ThyX-containing organisms, DHFR is absent. While the resulting influence on the folate metabolism of ThyX-containing bacteria is not fully understood, the presence of ThyX may provide growth benefits under conditions where the level of reduced folate derivatives is compromised. Interestingly, the third key enzyme implicated in generation of MTHF, serine hydroxymethyltransferase (SHMT), has a universal phylogenetic distribution, but remains understudied in ThyX-containg bacteria. To obtain functional insight into these ThyX-dependent thymidylate/folate cycles, we characterized the predicted SHMT from the ThyX-containing bacterium Helicobacter pylori. Serine hydroxymethyltransferase activity was confirmed by functional genetic complementation of a glyA-inactivated E. coli strain. A H. pylori ΔglyA strain was obtained, but exhibited markedly slowed growth and had lost the virulence factor CagA. Biochemical and spectroscopic evidence indicated formation of a characteristic enzyme-PLP-glycine-folate complex and revealed unexpectedly weak binding affinity of PLP. The three-dimensional structure of the H. pylori SHMT apoprotein was determined at 2.8Ǻ resolution, suggesting a structural basis for the low affinity of the enzyme for its cofactor. Stabilization of the proposed inactive configuration using small molecules has potential to provide a specific way for inhibiting HpSHMT.
Databáze: OpenAIRE
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