96 research outputs found
Preparation of the Peptide Containing the Essential Methionine Residue of Cytoplasmic Malate Dehydrogenase from Pig Heart
The essential methionine residue in the active site of cytoplasmic
malate dehydrogenase from pig heart has been labelled
with (14C)iodoacetate. The peptide, comprising this methionine
residue, has been isolated and purified. The amino acid analysis
showed that it is a hexapeptide of a composition: (Ser, Pro, Gly,
Val, Met)-Arg
Evidence for a histidine and a cysteine residue in the substrate-binding site of yeast alcohol dehydrogenase
Thermal inactivation and conformational lock studies on glucose oxidase
In this study, the dissociative thermal inactivation
and conformational lock theories are applied for the
homodimeric enzyme glucose oxidase (GOD) in order to
analyze its structure. For this purpose, the rate of activity
reduction of glucose oxidase is studied at various temperatures
using b-D-glucose as the substrate by incubation of
enzyme at various temperatures in the wide range between
40 and 70 �C using UV–Vis spectrophotometry. It was
observed that in the two ranges of temperatures, the
enzyme has two different forms. In relatively low temperatures,
the enzyme is in its dimeric state and has normal
activity. In high temperatures, the activity almost disappears
and it aggregates. The above achievements are confirmed
by dynamic light scattering. The experimental
parameter ‘‘n’’ as the obvious number of conformational
locks at the dimer interface of glucose oxidase is obtained
by kinetic data, and the value is near to two. To confirm the
above results, the X-ray crystallography structure of the
enzyme, GOD (pdb, 1gal), was also studied. The secondary
and tertiary structures of the enzyme to track the thermal
inactivation were studied by circular dichroism and
fluorescence spectroscopy, respectively. We proposed a
mechanism model for thermal inactivation of GOD based
on the absence of the monomeric form of the enzyme by
circular dichroism and fluorescence spectroscopy
Medium- and short-chain dehydrogenase/reductase gene and protein families: The MDR superfamily
The MDR superfamily with ~350-residue subunits contains the classical liver alcohol dehydrogenase (ADH), quinone reductase, leukotriene B4 dehydrogenase and many more forms. ADH is a dimeric zinc metalloprotein and occurs as five different classes in humans, resulting from gene duplications during vertebrate evolution, the first one traced to ~500 MYA (million years ago) from an ancestral formaldehyde dehydrogenase line. Like many duplications at that time, it correlates with enzymogenesis of new activities, contributing to conditions for emergence of vertebrate land life from osseous fish. The speed of changes correlates with function, as do differential evolutionary patterns in separate segments. Subsequent recognitions now define at least 40 human MDR members in the Uniprot database (corresponding to 25 genes when excluding close homologues), and in all species at least 10888 entries. Overall, variability is large, but like for many dehydrogenases, subdivided into constant and variable forms, corresponding to household and emerging enzyme activities, respectively. This review covers basic facts and describes eight large MDR families and nine smaller families. Combined, they have specific substrates in metabolic pathways, some with wide substrate specificity, and several with little known functions
Acetylcholinesterase Sensor Based on Polyelectrolyte Complexes with DNA Inclusion for the Determination of Reversible Inhibitors
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