Analysis of phosphofructokinase-1 activity as affected by pH and ATP concentration.
ATP
Enzyme activity
Initial-velocity method
Kinetic modeling
PFK1
Substrate inhibition
pH
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
11 Sep 2024
11 Sep 2024
Historique:
received:
29
02
2024
accepted:
03
09
2024
medline:
12
9
2024
pubmed:
12
9
2024
entrez:
11
9
2024
Statut:
epublish
Résumé
A key player in energy metabolism is phosphofructokinase-1 (PFK1) whose activity and behavior strongly influence glycolysis and thus have implications in many areas. In this research, PFK1 assays were performed to convert F6P and ATP into F-1,6-P and ADP for varied pH and ATP concentrations. PFK1 activity was assessed by evaluating F-1,6-P generation velocity in two ways: (1) directly calculating the time slope from the first two or more datapoints of measured product concentration (the initial-velocity method), and (2) by fitting all the datapoints with a differential equation explicitly representing the effects of ATP and pH (the modeling method). Similar general trends of inhibition were shown by both methods, but the former gives only a qualitative picture while the modeling method yields the degree of inhibition because the model can separate the two simultaneous roles of ATP as both a substrate of reaction and an inhibitor of PFK1. Analysis based on the model suggests that the ATP affinity is much greater to the PFK1 catalytic site than to the inhibitory site, but the inhibited ATP-PFK1-ATP complex is much slower than the uninhibited PFK1-ATP complex in product generation, leading to reduced overall reaction velocity when ATP concentration increases. The initial-velocity method is simple and useful for general observation of enzyme activity while the modeling method has advantages in quantifying the inhibition effects and providing insights into the process.
Identifiants
pubmed: 39261563
doi: 10.1038/s41598-024-72028-4
pii: 10.1038/s41598-024-72028-4
doi:
Substances chimiques
Adenosine Triphosphate
8L70Q75FXE
Phosphofructokinase-1
EC 2.7.1.11
Fructosephosphates
0
Adenosine Diphosphate
61D2G4IYVH
fructose-6-phosphate
6814-87-5
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
21192Subventions
Organisme : National Institute of Food and Agriculture
ID : 2017-67017-26470
Informations de copyright
© 2024. The Author(s).
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