Multiplex inhibitor screening and kinetic constant determinations for yeast hexokinase using mass spectrometry based assays
نویسندگان
چکیده
منابع مشابه
Kinetic studies of yeast hexokinase.
The mechanism of calf brain hexokinase has recently been studied in our laboratory (1). It was concluded from kinetic experiments that adenosine diphosphate (ADP) dissociates from the enzyme before addition of substrate glucose. This hypothesis was supported by kinetic studies of n-mannose, ADP, and glucose g-phosphate inhibition. The experiments appear to lend credence to theories in which an ...
متن کاملSolid phase capturable dideoxynucleotides for multiplex genotyping using mass spectrometry.
We report an approach using solid phase capturable biotinylated dideoxynucleotides (biotin-ddNTPs) in single base extension for multiplex genotyping by mass spectrometry (MS). In this method, oligonucleotide primers that have different molecular weights and that are specific to the polymorphic sites in the DNA template are extended with biotin-ddNTPs by DNA polymerase to generate 3'-biotinylate...
متن کاملYeast-based assays for screening 11β-HSD1 inhibitors.
BACKGROUND Intracellular metabolism of glucocorticoid hormones plays an important role in the pathogenesis of metabolic syndrome and regulates, among many physiological processes, collagen metabolism in skin. At the peripheral level the concentration of active glucocorticoids is mainly regulated by the 11β-hydroxysteroid dehydrogenase 1 (11β-HSD1) enzyme, involved in the conversion of cortisone...
متن کاملSubstrate synergism and the kinetic mechanism of yeast hexokinase.
Michaelis constants for MgATP with yeast hexokinase vary from 28 microM with D-mannose to above 4 mM for the slow ATPase reaction, with the different values reflecting the degree of synergism in binding of MgATP and the sugar substrate. The best substrates show the greatest synergism, but the correlation is not exact. Similar synergistic binding between MgADP or its methylene analogue and phosp...
متن کاملBioaffinity Mass Spectrometry Screening Using Droplet-Based Microfluidics
Bioaffinity mass spectrometry screening is a novel approach using non-denaturing electrospray ionization (ESI) mass spectrometry (MS) in identifying drug leads. This screening technique can detect and preserve noncovalent protein-active drug ligand complexes under different physiological conditions. Although there are many successful screening campaigns employing this technique, the big challen...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: Journal of the American Society for Mass Spectrometry
سال: 2003
ISSN: 1044-0305,1879-1123
DOI: 10.1016/s1044-0305(02)00867-x