[Physico-chemical properties of thiamine kinase from Saccharomyces carlsbergensis yeasts].
The amino acid composition and intrinsic fluorescence were studied in thiamine kinase (ES 2.7.6.2) of brewer's yeast. The enzyme molecule is characterized by higher concentrations of amino acids which promote alpha-helix formation of the protein globule, the amount of residues (cysteine, proline) either binding or folding polypeptide chains being considerably high. Amino acids of middle and low hydrophobicities were the most frequent among the amino acid residues with nonpolar R-groups. The value for the protein isoelectric point was 6.21. The eigen pH value and isoionic point were in good agreement with the isoelectric point value and amounted to 6.28. The fluorescence spectrum has a maximum at 328 nm, half-width at 53 nm and a quantum yield at 0.14 nm. The tryptophane residues were located in hydrophobic surroundings, unexposed to anion quenchers and almost unexposed to cation ones. The fluorescence and phosphofluorescence parameters were sensitive to the conformational changes in the molecule. At pH of 5-9 the protein conformation remained unchanged. The temperature rise above 40 degrees C resulted in a disturbance in the nativity of the globule. The elevation of the enzyme concentration from 0.05 to 1 mg/ml increased the polarization degree from 0.115 to 0.194, the quantum yield and the spectrum position remaining unchanged. The results obtained develop knowledge of the equilibrium system of oligomerous forms of thiamine kinase with different catalytic properties.
Chernikevich
I P
IP
Maskevich
A A
AA
Naumov
A V
AV
Maskevich
S A
SA
Kivach
L N
LN
Voskoboev
A I
AI
D000596:Amino Acids / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D010770:Phosphotransferases / Q000378:metabolism*; D017853:Phosphotransferases (Alcohol Group Acceptor); D011487:Protein Conformation; D012440:Saccharomyces / Q000201:enzymology*; D013050:Spectrometry, Fluorescence
D000596:Amino Acids; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C035182:thiamin kinase
Fiziko-khimicheskie svoĭstva tiaminkinazy drozhzheĭ Saccharomyces carlsbergensis.