ABSTRACT
New microcarriers for the growth of animal cells have been synthesized and studied. The preparations are porous cellulose beads, modified by diamines. Spreading and growth of L cells, MEVO and HETR cells on these beads were observed. As a result of the cultivation the number of animal cells increased 5-10-fold.
Subject(s)
Cells, Cultured/cytology , Animals , Cell Adhesion , Cell Division , Cellulose , Cytological Techniques/instrumentation , Diamines/pharmacology , LigandsABSTRACT
Two highly purified forms of acid-stable proteinase inhibitor from urine of pregnant women with nephropathies (Mr 22000 and 32000, ASI-22 and ASI-32, respectively) were obtained. The preparations were homogenous in molecular mass and polymorphous in molecular charge (pI from 3.9 to 4.2). ASI-22 an ASI-32 effectively inhibited trypsin and chymotrypsin (Ki approximately 1 x 10(-8)-1 x 10(-9) M, ka approximately 10(5)M-1 sec-1, kd approximately 3 x 10(-4) sec-1) by the permanent mechanism of action. Both forms inhibited the esterase activity of pancreatic pig elastase by the progressive mechanism of action (ki approximately 1 x 10(4)M-1 min-1 at 37 degrees). Rabbit monospecific antiserum to total ASI-22 and ASI-32 preparation was obtained 1 ml of the anti-ASI-serum contained 95 micrograms of antibodies. Total ASI preparations was immunochemically homogenous and had antigenic similarity to inter-alpha-trypsin inhibitor from human plasma.
Subject(s)
Immune Sera/isolation & purification , Pregnancy Complications/urine , Protease Inhibitors/urine , Animals , Chymotrypsin/antagonists & inhibitors , Drug Stability , Female , Humans , Kidney Diseases/immunology , Kidney Diseases/urine , Methods , Molecular Weight , Pregnancy , Pregnancy Complications/immunology , Pregnancy Trimester, Third , Protease Inhibitors/immunology , Protease Inhibitors/isolation & purification , Rabbits , Trypsin Inhibitors/pharmacologyABSTRACT
The substrate specificity of acylase I from pig kidney has been studied. For the N-acetyl derivatives of D,L-amino acids with an unbranched side chain the catalytic efficiency of enzymatic hydrolysis increases linearly with an increase of the substrate hydrophobicity. The tangent of the linear dependence of logarithm of the second order rate constant of enzymatic hydrolysis on the Hansch hydrophobicity constant is 0,7. This supports a simple "extraction" model of energy realization during the enzyme binding to the substrate in the course of the reaction. When the amino acids with a branched side chain are used, the efficiency of hydrolysis of N-acetyl derivatives decreases by one order of magnitude as compared to the amino acids with an unbranched side chain having the same number of carbon atoms. The N-acetyl derivatives of aromatic amino acids are hydrolyzed in a small degree. An analysis of the substrate specificity of pig kidney acylase I demonstrates that the enzyme has a narrow hydrophobic "cleft" with the length not less than four metrylene links.
