BIOCHEMICAL PROPERTIES OF CARBOXYPEPTIDASE A FROM NON-TRANSFERRED TISSUE AND BENIGNANT TUMOR TISSUE OF THE MAMMALIAN GLAND
Journal Title: Вісник Одеського національного університету. Біологія - Year 2016, Vol 21, Issue 2
Abstract
Introduction. Lysosomal carboxypeptidase A [КE 3.4.17.1] is a zinc-dependent protease that participates actively at the terminal stages of proteolysis and regulation of metabolism of cells in many physiological and pathological processes. Purpose. To study biochemical properties of carboxypeptidase A from non-transformed tissue and benignant tumor tissue of the mammalian gland. Methods. Sampling of anatomical materials for research was conducted with compliance of ethical and legal standards. Excretion of the enzymes included gradual fractionation with the (NH4)2SO4, dialysis in presence of 2.0 мМ Zn++ and gel chromatography on the sephadex – G 75. The investigation of the substrate specificity of the enzymes was held by hydrolysis of the substrate of carbobenzoxyphenylalanine, phenylalanylalanine, glutamyltyrosine, prolylalanine (2.0 mM), haemoglobin and casein (2.0 %). The influence of inhibitors and activators was determined in presence of: DTT, Zn++, cysteine, triton X-100, soybean trypsin inhibitor, leupeptin, pepstatin, PHMB, FMSF, dimethylmolyemydanhydride, tozylheptanol, mercaptoethanol, EDTA and 1.10 - phenanthroline. The maximal velocity (Vmax), Mihaelis constant (Km), inhibition type and inhibition constant (Ki) analysed by Laynuiveru – Berku method. Results. Carboxypeptidase A from non-transformed tissue and benignant tumor of the mammalian gland better splits the substrates, which have hydrophobic and aromatic amino acids. The activity of carboxypeptidase A from the benignant tumor of the mammalian gland is inhibited most of all under influence of leupeptin, tozylheptanol and dimethylmolyemydanhydride, in contrast to non-transformed tissue. For carboxypeptidase A from non-transformed tissue of the mammalian gland Km = 0.24 mM and Ki = 0.40 mM were determined, for carboxypeptidase A from the benignant tumor tissue of the mammalian gland – Km = 0.14 mM and Ki = 0.16 mM. Conclusion. Carboxypeptidase A from non-transformed tissue and benignant tumor of the mammalian gland is identical as to the substrate specificity, inhibition by phenylalanine for noncompetitive type, inhibition and activation effect by reagent with the exclusion of leupeptin, tozylheptanol and dimethylmolyemydanhydride, but differ as to affinity to carbobenzoxyphenylalanine and sensitivity to phenylalanine.
Authors and Affiliations
K. A. Filiptsova, I. L. Vovchuk
Keywords
Related Articles
- EP ID EP308988
- DOI -
- Views 78
- Downloads 0
GENDER DIFFERENCES OF LIQUOR BRAIN SYSTEM IN PERSONS OF ALL AGES
Tomogramms analyzed 69 individuals of all ages without the expressed pathological changes in the brain using morphometric methods according to the recommendations of encephalometry. There were studied 13 morphometric par...
ASSESSMENT OF TOXICITY AND POLLUTION OF BOTTOM SEDIMENTS IN THE NORTHWESTERN PART OF THE BLACK SEA WITH HEAVY METALS
The northwestern shelf is the most polluted regions of Black sea. The aim of the work was to compare the levels of bottom sediment’s toxicity and pollution with heavy metals (As, Cd, Hg, Pb, Zn, Cu, Ni, Co, Cr) in the Od...
EVALUATION OF REPRESENTATIVENESS OF THE NATURAL RESERVE FUND OF ODESA REGION FOR PROTECTION OF ECOSYSTEMS AND SOZOPHITES IN THE CONTEXT OF GEOBOTHANICAL ZONING
Geobotanical zoning of Odesa Region was clarified. 14 geobotanical areas were allocated within 6 districts. We have considered the distribution of objects of Natural Reserve Fund by areas and districts. It has been revea...
DENDROFLORA OF THE IZMAIL FORTRESS (IZMAIL, ODESSA REGION)
The research was carried out by traditional methods. Species names and family volumes are by Vascular plants of Ukraine checklist (Mosyakin, Fedoronchuk, 1999), life formes are by I. G. Serebryakov (1962), areas of speci...
ANALYSIS OF THE SOZOPHYTES OF THE CHYGRYNSKYI RAVINE (ODESA REGION, UKRAINE)
Steppe biome is practically destroyed in Ukraine. Therefore, areas with steppe plants should be taken under special protection. During 2013-2016 the authors studied flora in Chygrynskyi Ravine, which covers an area of ab...