Relationships between kinetic constants and the amino acid composition of enzymes from the yeast Saccharomyces cerevisiae glycolysis pathway.

Abstract

The kinetic models of metabolic pathways represent a system of biochemical reactions in terms of metabolic fluxes and enzyme kinetics. Therefore, the apparent differences of metabolic fluxes might reflect distinctive kinetic characteristics, as well as sequence-dependent properties of the employed enzymes. This study aims to examine possible linkages between kinetic constants and the amino acid (AA) composition (AAC) for enzymes from the yeast Saccharomyces cerevisiae glycolytic pathway. The values of Michaelis-Menten constant (KM), turnover number (kcat), and specificity constant (ksp = kcat/KM) were taken from BRENDA (15, 17, and 16 values, respectively) and protein sequences of nine enzymes (HXK, GADH, PGK, PGM, ENO, PK, PDC, TIM, and PYC) from UniProtKB. The AAC and sequence properties were computed by ExPASy/ProtParam tool and data processed by conventional methods of multivariate statistics. Multiple linear regressions were found between the log-values of kcat (3 models, 85.74% < Radj.2 <94.11%, p < 0.00001), KM (1 model, Radj.2 = 96.70%, p < 0.00001), ksp (3 models, 96.15% < Radj.2 < 96.50%, p < 0.00001), and the sets of AA frequencies (four to six for each model) selected from enzyme sequences while assessing the potential multicollinearity between variables. It was also found that the selection of independent variables in multiple regression models may reflect certain advantages for definite AA physicochemical and structural propensities, which could affect the properties of sequences. The results support the view on the actual interdependence of catalytic, binding, and structural residues to ensure the efficiency of biocatalysts, since the kinetic constants of the yeast enzymes appear as closely related to the overall AAC of sequences.