Structural and biochemical analysis of phosphoethanolamine methyltransferase from the pine wilt nematode Bursaphelenchus xylophilus

Autor: Kaisen Yao, Divya Rayapati, Janki K. Patel, Andreia Silva da Rocha, Alex J. Petronio, Grant B. Nelson, Angela J. Zou, John A. Rincon, Mythili Ramachandran, Soon Goo Lee, Kevin Kulshrestha, Margery Gang, Melanie A. Sparks, Peter R. Qin, Micah K. Steinbrecher, Raymundo Marcelo, Barrie Cascella, Joan Gabriel Rodinho Nunes Ferreira, Wilhelm Cruz, Cherry Jiang, Vidhya M. Meyyappa, Eric J. Zhang, Jakob H.P. Kroboth, Joseph M. Jez, Rohit V. Jaswaney, Samantha K. Powers, Antea J. DeMarsilis, Cynthia K. Holland, Michelle S. Chung
Rok vydání: 2020
Předmět:
Models
Molecular
Protein Conformation
alpha-Helical
Methyltransferase
Nematoda
030231 tropical medicine
Genetic Vectors
Gene Expression
Bursaphelenchus xylophilus
Biology
Crystallography
X-Ray
Substrate Specificity
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
Protein structure
Escherichia coli
Animals
Protein Interaction Domains and Motifs
Amino Acid Sequence
Binding site
Cloning
Molecular
Molecular Biology
030304 developmental biology
Phosphocholine
Plant Diseases
0303 health sciences
Binding Sites
Sequence Homology
Amino Acid
Mutagenesis
Active site
Methylation
Helminth Proteins
Methyltransferases
biology.organism_classification
Pinus
Recombinant Proteins
Kinetics
Biochemistry
chemistry
Ethanolamines
biology.protein
Thermodynamics
Parasitology
Protein Conformation
beta-Strand
Sequence Alignment
Protein Binding
Zdroj: Molecular and biochemical parasitology. 238
ISSN: 1872-9428
Popis: In free-living and parasitic nematodes, the methylation of phosphoethanolamine to phosphocholine provides a key metabolite to sustain phospholipid biosynthesis for growth and development. Because the phosphoethanolamine methyltransferases (PMT) of nematodes are essential for normal growth and development, these enzymes are potential targets of inhibitor design. The pine wilt nematode (Bursaphelenchus xylophilus) causes extensive damage to trees used for lumber and paper in Asia. As a first step toward testing BxPMT1 as a potential nematicide target, we determined the 2.05 A resolution x-ray crystal structure of the enzyme as a dead-end complex with phosphoethanolamine and S-adenosylhomocysteine. The three-dimensional structure of BxPMT1 served as a template for site-directed mutagenesis to probe the contribution of active site residues to catalysis and phosphoethanolamine binding using steady-state kinetic analysis. Biochemical analysis of the mutants identifies key residues on the β1d-α6 loop (W123F, M126I, and Y127F) and β1e-α7 loop (S155A, S160A, H170A, T178V, and Y180F) that form the phosphobase binding site and suggest that Tyr127 facilitates the methylation reaction in BxPMT1.
Databáze: OpenAIRE
Externí odkaz: