Alkaline phosphatase leucine

Isoenzymes or isozymes are enzymes from a single species that have the same kind of enzymatic activity but differ in chemical structure. In addition, they may differ in quantitative characteristics such as possessing different Km s with the same substrate and may differ in response to temperature and effectors. Isozymes of more than 100 enzymes have been demonstrated in humans. The most important of these for diagnostic purposes are the isozymes of LDH, CK, alkaline phosphatase, leucine aminopeptidase, acid phosphatase, and aldolase. These have been exploited for differential organ diagnosis. 

The term enzymatic markers of cholestasis (or cholestasis-indicating enzymes) comprises alkaline phosphatase, leucine aminopeptidase (or leucine arylamidase) as well as 5 -nucleotidase and y-glutamyl transferase, (see chapter 13)... 

Proximal tubule cells in culture should have retained functional attributes such as (1) polar architecture and junctional assembly of epithelia and correct membrane distribution of enzymes and transport systems (2) vectorial transport of solutes and water, manifested by the formation of domes when cultured on solid supports [81] and the generation of transepithelial electrophysiological properties [82, 83] due to the expression of proximal tubule specific claudins 2- and 10 [84, 85] (3) cellular uptake of xenobiotics from either the apical or basolateral side, as observed in vivo and (4) expression of nephron segment-specific characteristics, i.e., distinct expression of differentiation markers, metabolic and transport properties, and hormone responsiveness. Such markers include the expression of the brush border enzymes alkaline phosphatase, leucine aminopeptidase, and y-glutamyl transferase [4, 86], In addition, proximal tubule cells should possess Na+,K+-ATPase activities, Na+-dependent glucose, and p-aminohippurate transport. Proximal tubule cells increase cAMP levels in response to parathyroid... 

Metabolic Functions. Zinc is essential for the function of many enzymes, either in the active site, ie, as a nondialyzable component, of numerous metahoenzymes or as a dialyzable activator in various other enzyme systems (91,92). WeU-characterized zinc metahoenzymes are the carboxypeptidases A and B, thermolysin, neutral protease, leucine amino peptidase, carbonic anhydrase, alkaline phosphatase, aldolase (yeast), alcohol... 

The enzymes found in liver cells (Group I enzymes) include more than a dozen enzymes used in diagnostic laboratories, but those used most commonly are the transaminases (GOT and GPT), which continue to be the most widely used indicators of liver cell integrity. Enzymes found in the biliary cells (Group II) include alkaline phosphatase, glutamyl-transferase, leucine amniopeptidase and 3-nucleotidase. 

Enzymes activities are particularly sensitive to the anticoagulant used in collecting the specimen. Heparin inhibits acid phosphatase (W16) and muramidase (Z5). Amylase activity is inhibited by oxalate or citrate (MIO), and lactic dehydrogenase and acid phosphatase lose activity in oxalate (C2). Alkaline phosphatase is stable in oxalate, oxalate-fluoride, or heparin, but 25 mAf citrate inhibits 50% of the activity, and as little as 50 mlf EDTA is completely inhibitory (B19). Leucine aminopeptidase is inhibited by EDTA, as is creatine phosphokinase (F3). Amylase activity has been reported to be only 83% of that in serum when oxalate or citrate-plasma is used (MIO). Heparin plasma appears to have no inhibitory effect. Despite the fact that clotting factor V is not stable in oxalate or EDTA, these are often used as anticoagulants to obtain plasma for prothrombin determinations (Z2, Z4). 

We shall now briefly outline some of the features of the zinc metalloenzymes which have attracted most research effort several reviews are available, these are indicated under the particular enzyme, and for more detailed information the reader is referred to these. Attention is focussed here, albeit briefly, on carbonic anhydrases,1241,1262,1268 carboxypeptidases, leucine amino peptidase,1241,1262 alkaline phosphatases and the RNA and DNA polymerases.1241,1262,1462 Finally, we examine alcohol dehydrogenases in rather more detail to illustrate the use of the many elegant techniques now available. These enzymes have also attracted much effort from modellers of the enzymic reaction and such studies, which reveal much interesting coordination chemistry and often new catalytic properties in their own right—and often little about the enzyme system itself (except to indicate possibilities), will be mentioned in the next section of this chapter. 

EDTA, probably by chelation of metallic cofactors, inhibits alkaline phosphatase, creatine kinase, and leucine aminopeptidase activities. Because it chelates calcium and iron, EDTA is unsuitable for specimens for calcium and iron analyses using photometric or titrimetric techniques. As an... 

Koleva M (1977) Changes in the urinary excretion of gamma-glutamyltranspeptidase, leucine aminopeptidase and alkaline phosphatase in the combined action of ethylene glycol and high temperature. Probl Khig 3 35-46... 

Measurement of serum y-GT activity has clinical significance. The enzyme is present in all tissues, but the highest level is in the kidney however, the serum enzyme originates primarily from the hepatobiliary system. Elevated levels of serum y-GT are found in the following disorders intra- and posthepatic biliary obstruction (elevated serum y-GT indicates cholestasis, as do leucine aminopeptidase, 5 -nucleotidase, and alkaline phosphatase) primary or disseminated neoplasms some pancreatic cancers, especially when associated with hepatobiliary obstruction alcohol-induced liver disease (serum y-GT may be exquisitely sensitive to alcohol-induced liver injury) and some prostatic carcinomas (serum from normal males has 50% higher activity than that of females). Increased activity is also found in patients receiving phenobarbital or phenytoin, possibly due to induction of y-GT in liver cells by these drugs. 

K23. Kowlessar, O. D., Haeffner, L. J., and Riley, E. M., Localization of serum leucine aminopeptidase, 5-nucleotidase and nonspecific alkaline phosphatase by starch-gel electrophoresis. Clinical and biochemical significance in disease states. Ann. N.Y. Acad. Sci. 94, 836-843 (1961). 

The nephrotoxicity of 16 continues to generate considerable interest. Orellanine was highly toxic to mice (LD50 = 12.5 mg/kg i.p.)[101] and caused interstitial nephritis and tubular necrosis in mouse kidney [102], A summary of 16-induced changes in renal function and morphology has been reported [103]. In LLC-PKi renal epithelial cell cultures, 16 decreased the activity of alkaline phosphatase and lactate dehydrogenase, and decreased the incorporation of H-leucine and H-thymidine [104]. Orellanine was a noncompetitive inhibitor of renal alkaline phosphatase, but a competitive inhibitor of the intestinal and placental enzymes [105]. In canine kidney MDCK cell cultures, 16, or a metabolite of 16, inhibited protein, RNA and DNA synthesis [106]. 

The reader is referred to the following sources of further information on the specific cocatalytic enzymes superoxide dismutase, and the reduced form, alkaline phosphatases, nuclease PI, purple acid phosphatase, amidohydrolase, leucine aminopeptidase, general comments on the mechanisms of the phosphatases and aminopeptidases, and other cocatalytic zinc enzymes. ... 

We shall now briefly outline some of the features of the zinc metalloenzymes which have attracted most research effort several reviews are available, these are indicated under the particular enzyme, and for more detailed information the reader is referred to these. Attention is focussed here, albeit briefly, on carbonic anhydrases, carboxypeptidases, leucine amino peptidase,alkaline phosphatases and the RNA and DNA polymerases. 

Brush-border membrane alkaline phosphatase (EC 3.1.3.1) of the rat small intestine was inactivated in an in vitro Fe /ascorbate oxygen-radical generating system (Dudeja and Brasitus 1993). The activities of sucrase, maltase, leucine amino-peptidase, and y-glutamyl transpeptidase were unchanged. 

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