Cobalt (3) carboxypeptidase A: preparation and esterase activity

Co(II) carboxypeptidase A has been oxidized to Co(III) carboxypeptidase A with hydrogen peroxide. The resultant metalloprotein has an absorption spectrum different from that of the Co(II) enzyme and the metal is no longer removable by dialysis. The Co(III) carboxypeptidase A retains esterase activity comparable to that of the Co(II) enzyme and has very low peptidase activity. This demonstrates that scission of a bond to the first coordination sphere of the metal is not necessary for the hydrolysis of ester substrates.     

Effect of testosterone on carboxypeptidase H activity in the murine hypothalamus and hypophysis

Effects of single intraperitoneal administration of testosterone propionate (3 or 30 mg/kg body weight) on activity of carboxypeptidase H in pituitary body and hypothalamus of female white mouse were studied. It was found that testosterone propionate treatment (3 and 30 mg/kg body weight) increased the carboxypeptidase H activity in pituitary through 0.5 hour and it decreased one in 24 hours after treatment. The carboxypeptidase H activity in hypothalamus was lower as compared with control animals in 24 hours after testosterone propionate treatment in the dose 3 mg/kg body weight. However, the carboxypeptidase H activity in hypothalamus was lower in 0.5 and 24 hours and it was higher in 4 h after testosterone propionate treatment in the dose 30 mg/kg body weight as compared with the control. These data suggest that testosterone affects the carboxypeptidase H activity by changing the level of enzyme gene expression.     

Effect of testosterone and progesterone on carboxypeptidase H activity during stress in mice

It was found that testosterone propionate (3 mg/kg of body weight) and progesterone (1 mg/kg of body weight) partially prevent an augmentation of the carboxypeptidase H activity in the mouse pituitary gland under stress caused by a single intraperitoneal administration of olive oil. In testicles, testosterone prevented augmentation of the enzyme activity within 0.5 hours but increased it through in 4 and 24 hours after treatment. Progesterone in testicles augmented the enzyme activity within 0.5 hours, but not in 4 and 24 hours. Sex steroids were not affected by stressinduced carboxypeptidase H activity in hypothalamus and adrenal medulla. It was concluded that carboxypeptidase H did not take part in integration of hypothalamus-pituitary-adrenal and hypothalamus-pituitary-gonadal axis under the stress.     

Effect of stress factors on carboxypeptidase H activity in areas of the rat brain]

It is established that carboxypeptidase H activity increases as affected by various stress factors in the rat brain departments. The increase of the enzyme activity because of the emotional-pain stress is of continuous character. Possible role of carboxypeptidase H in the development of the stress response is discussed.     

Effect of chronic ethanol consumption on basic carboxypeptidase activity in regions of the rat brain

It is discovered that chronic consumption of ethanol induced decrease of carboxypeptidase H activity in striatum by 27%; increase of carboxypeptidase M activity in hippocampus by 67% and decrease in cerebral hemispheres by 34%; phenylmethylsulfonyl fluoride-inhibited carboxypeptidase activity increase in hypothalamus by 141%, in striatum by 60% and in optic and lamina quadrigemina by 34%. The role of basic carboxypeptidases in mechanisms of ethanol influence on the peptidergic systems are discussed.     

1,2-二亚油酸-3-硬脂酸-甘油三酯对玉米象酯酶及其同工酶活性的影响

研究葫芦巴(Trigonella foenum graecumL.)种子提取物1,2-二亚油酸-3-硬脂酸-甘油三酯对玉米象成虫酯酶的影响。结果表明,玉米象成虫被1,2-二亚油酸-3-硬脂酸-甘油三酯触杀处理后,8h之内,虫体内酯酶比活力呈下降趋势,8h后开始上升,24h达到最大值,48h又有所下降。酯酶同工酶电泳结果显示,处理后的玉米象的酯酶同工酶中,增加了1条迁移率较慢的条带和1条迁移率较快的条带。     

Effect of atherosclerosis on lysosomal cholesterol esterase activity in rabbit aorta.

Radiolabeled cholesteryl oleate, when incorporated into phospholipid vesicles, was hydrolyzed at acid pH by an enzyme present in rabbit aortic homogenates. In contrast, cholesteryl oleate presented as an acetone dispersion was not effectively hydrolyzed at acid pH under identical conditions. Using the vesicle preparation as substrate, a sensitive assay system for the acid hydrolase was developed in which hydrolysis was proportional to protein concentration and incubation time, and was independent of substrate concentration. The physical state of the vesicles was apparently not altered by the assay conditions, and no hydrolysis of the vesicle-associated phospholipid was detected. Acid cholesterol esterase activity in atherosclerotic aortic tissue was 2.5-fold greater than that of control tissue, and even greater increases were observed in the activities of other lysosomal enzymes (N-acetyl-beta-d-glucosaminidase and beta-glucuronidase). Glucose-6-phosphatase activity was also increased in aortas from cholesterol-fed animals while 5' nucleotidase activity remained unchanged. Labeled triolein also was incorporated into phospholipid vesicles and was hydrolyzed by an acid lipase in aortic tissue. Similarities between triolein and cholesteryl oleate hydrolysis existed with respect to pH optimum and the effect of cholesterol feeding on activity, suggesting that a single enzyme may hydrolyze both lipids.     

A rapid spectrophotometric method for the determination of esterase activity

We have developed a spectrophotometric assay method which continuously records esterase activity at 510 nm by monitoring absorbance changes due to the formation of a diazo dye complex. In our method, α-naphthyl ester substrates are hydrolyzed by enzymatic action to α-naphthol which couples to Fast Blue RR salt (a diazonium salt) forming a diazo dye complex. Our method is unique in directly monitoring the formation of the diazo dye complex without extracting the color of the complex as in other methods that use naphthyl esters and diazo coupling of reaction products. The method appears to be limited to α-naphthyl ester substrates, however, since β-naphthyl esters did not give a linear change in absorbance in the enzymatic reactions tested. With this assay method, one can use a single substrate both to determine esterase units quantitatively in solution and to detect esterase staining activity on gel electrophoresis.     

A unique activity assay for carboxypeptidase A in human serum

Measurement of carboxypeptidase A, one of the pancreatic proteolytic enzymes, in human serum is made possible by a combination of affinity chromatography to isolate and concentrate the enzyme followed by monitoring activity spectrophotometrically with a high-turnover peptide substrate. Concentrations of enzyme in the nanogram-per-milliliter range can be determined with high precision and reliability. Initial clinical application of this method demonstrates no detectable activity in serum from normal individuals, but the enzyme is present in the sera of individuals with pancreatitis.     

Catalytic activity of carboxypeptidase B and of carboxypeptidase Y with anisylazoformyl substrates

Anisylazoformyllysine (CH3OC6H4-N = N-CO-Lys-OH) is rapidly hydrolyzed at the acyl-lysine linkage by the zinc-enzyme porcine carboxypeptidase B. The catalytic reaction is readily monitored spectrophotometrically by disappearance of the intense absorption (348.5 nm, epsilon 18400) of the azo chromophore, which chemically fragments after substrate cleavage. Carboxypeptidase Y has no activity toward this type of substrate.