Kinetic analysis of PRMT1 reveals multifactorial processivity and a sequential ordered mechanism

Autor: Brown, Jennifer I, Koopmans, Timo, van Strien, Jolinde, Martin, Nathaniel I, Frankel, Adam, Afd Chemical Biology and Drug Discovery, Chemical Biology and Drug Discovery
Přispěvatelé: Afd Chemical Biology and Drug Discovery, Chemical Biology and Drug Discovery
Jazyk: angličtina
Rok vydání: 2017
Předmět:
0301 basic medicine
Protein-Arginine N-Methyltransferases
Arginine
Biology
Biochemistry
Methylation
Mass Spectrometry
Substrate Specificity
03 medical and health sciences
0302 clinical medicine
Arginine/metabolism
Protein-Arginine N-Methyltransferases/genetics
Genetics
medicine
Humans
Enzyme kinetics
Amino Acid Sequence
Recombinant Proteins/biosynthesis
Molecular Biology
030304 developmental biology
chemistry.chemical_classification
0303 health sciences
Repressor Proteins/genetics
030102 biochemistry & molecular biology
Organic Chemistry
Substrate (chemistry)
Processivity
Peptides/analysis
Recombinant Proteins
Repressor Proteins
Kinetics
030104 developmental biology
Enzyme
Mechanism of action
chemistry
Biophysics
Biocatalysis
Molecular Medicine
medicine.symptom
Peptides
030217 neurology & neurosurgery
Function (biology)
Biotechnology
Zdroj: ChemBioChem, 19(1), 85-99
ChemBioChem
ChemBioChem, 19(1), 85. Wiley-VCH Verlag
ISSN: 1439-4227
Popis: Arginine methylation is a prevalent post‐translational modification in eukaryotic cells. Two significant debates exist within the field: do these enzymes dimethylate their substrates in a processive or distributive manner, and do these enzymes operate using a random or sequential method of bisubstrate binding? We revealed that human protein arginine N‐methyltransferase 1 (PRMT1) enzyme kinetics are dependent on substrate sequence. Further, peptides containing an Nη‐hydroxyarginine generally demonstrated substrate inhibition and had improved KM values, which evoked a possible role in inhibitor design. We also revealed that the perceived degree of enzyme processivity is a function of both cofactor and enzyme concentration, suggesting that previous conclusions about PRMT sequential methyl transfer mechanisms require reassessment. Finally, we demonstrated a sequential ordered Bi–Bi kinetic mechanism for PRMT1, based on steady‐state kinetic analysis. Together, our data indicate a PRMT1 mechanism of action and processivity that might also extend to other functionally and structurally conserved PRMTs.
Databáze: OpenAIRE
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