Carbonic anhydrase mechanisms

Structure and physiology of the kidney glomerular filtration tubular activity selective reabsorption and secretion, often using specific carrier mechanisms carbonic anhydrase and acid-base balance. The kidney also produces, and is sensitive to, hormones actions of the hormones ADH, aldosterone and PTH the kidney as a secretory organ erythropoietin, the renin-angiotensin system vitamin D3. 

Theoretical calculations have been carried out on a number of zinc-containing enzymatic systems. For example, calculations on the mechanism of the Cu/Zn enzyme show the importance of the full protein environment to get an accurate description of the copper redox process, i.e., including the electronic effects of the zinc ion.989 Transition structures at the active site of carbonic anhydrase have been the subject of ab initio calculations, in particular [ZnOHC02]+, [ZnHC03H20]+, and [Zn(NH3)3HC03]+.990... 

Thermal reaction techniques enable a quantification of the influence of solvation on reactivities.1,2,19 One particular reaction which is a good example of how solvation can affect the nature of a core ion reaction site comes from a study118 of the interaction of OH with C02. The gas-phase reaction between the individual species is quite exothermic and can only take place by a three-body association mechanism. The reaction proceeds very slowly in the liquid phase and has been calculated119 to have a barrier of about 13 kcal mol-1. In biological systems, the reaction rate is enhanced by about 4 orders of magnitude through the enzyme carbonic anhydrase. Recent studies carried out in our laboratory provide detailed... 

Certain enzymes shown to be present in myelin could be involved in ion transport. Carbonic anhydrase has generally been considered a soluble enzyme and a glial marker but myelin accounts for a large part of the membrane-bound form in brain. This enzyme may play a role in removal of carbonic acid from metabolically active axons. The enzymes 5 -nucleotidase and Na+, K+-ATPase have long been considered specific markers for plasma membranes and are found in myelin at low levels. The 5 -nucleotidase activity may be related to a transport mechanism for adenosine, and Na+, K+-ATPase could well be involved in transport of monovalent cations. The presence of these enzymes suggests that myelin may have an active role in ion transport in and out of the axon. In connection with this hypothesis, it is of interest that the PLP gene family may have evolved from a pore-forming polypeptide [9],... 

The identification of different carbonate binding modes in copper(II) and in zinc(II)/2,2 -bipyridine or tris(2-aminoethyl)amine/(bi)carbonate systems, specifically the characterization by X-ray diffraction techniques of both r)1 and r 2 isomers of [Cu(phen)2(HC03)]+ in their respective perchlorate salts, supports theories of the mechanism of action of carbonic anhydrase which invoke intramolecular proton transfer and thus participation by r)1 and by r 2 bicarbonate (55,318). 

The most fundamental process dealing with the activation of C02 involves the hydration of C02 to produce bicarbonate and the reverse dehydration of bicarbonate to produce C02. These processes are of biological and environmental significance since they control the transport and equilibrium behavior of C02. The spontaneous hydration of C02 and dehydration of HCO3 are processes that are too slow and must therefore be catalyzed by metal complexes in order to expedite the overall conversion rate. In biological systems, a series of enzymes, the carbonic anhydrases, are the efficient catalysts and can accelerate the reactions by up to 7 orders of magnitude. The mechanism of this... 

The first zinc enzyme to be discovered was carbonic anhydrase in 1940, followed by car-boxypeptidase A some 14 years later. They both represent the archetype of mono-zinc enzymes, with a central catalytically active Zn2+ atom bound to three protein ligands, and the fourth site occupied by a water molecule. Yet, despite the overall similarity of catalytic zinc sites with regard to their common tetrahedral [(XYZ)Zn2+-OH2] structure, these mononuclear zinc enzymes catalyse a wide variety of reactions, as pointed out above. The mechanism of action of the majority of zinc enzymes centres around the zinc-bound water molecule,... 

Figure 12.3 (a) The active site of human carbonic anhydrase and (b) a simplified mechanism of action for the enzyme B = general base, probably His64. (Reprinted with permission from Parkin, 2004. Copyright (2004) American Chemical Society.)... 

Menziani, M.C., De Benedetti, P.G., Gago, F. and Richards, W.G. (1989) The binding of benzenesulfonamides to carbonic anhydrase enzyme. A molecular mechanics study and quantitative structure-activity... 

Part of the metabolic machinery of an osteoclast resembles the red cell and the renal tubule cells because all of these cell types contain the enzyme carbonic anhydrase (carbonate dehydratase) which generates acid, that is protons, and have ion pumps in their plasma membranes. The mechanism of bone resorption requires the action of cathepsin and metalloproteinase-9 working in an acidic environment (Figure 9.8). 

C. K. Tu, D. N. Silverman, C. Forsman, B. H. Jonsson, S. Lindskog, Role of Histidine 64 in the Catalytic Mechanism of Human Carbonic Anhydrase II Studied with a Site-Specific Mutant , Biochemistry 1989, 28, 7913-7918. 

The carbonic anhydrase (Cam) in M. thermophila cells is elevated several fold when the energy source is shifted to acetate, suggesting a role for this enzyme in the acetate-fermentation pathway. It is proposed that Cam functions outside the cell membrane to convert CO2 to a charged species (reaction A4) thereby facilitating removal of product from the cytoplasm. Cam is the prototype of a new class (y) of carbonic anhydrases, independently evolved from the other two classes (a and P). The crystal structure of Cam reveals a novel left-handed parallel P-helix fold (Kisker et al. 1996). Apart from the histidines ligating zinc, the activesite residues of Cam have no recognizable analogs in the active sites of the a- and P-classes. Kinetic analyses establish that the enzyme has a zinc-hydroxide mechanism similar to that of Cab (Alber et al. 1999). 

Drugs of this group inhibit activity of carbonic anhydrase, an enzyme that catalyzes the reversible reaction of water and carbon dioxide, which forms carbonic acid. The mechanism of action of this group of drags is not fuUy understood. However, inhibition of carbonic anhydrase activity leads to a reduction of carbonic acid formation and an increase in bicarbonate, sodium, and potassium excretion with urine, which eventually leads to a significant increase in the process of excreting water from the organism. 

Fig. 23. A general mechanism of CO2 hydration as catalyzed by carbonic anhydrase II. Certain structural details (e.g., the function of pentacoordinate zinc or the degree of CO2—Zn interaction in enzyme-substrate association) remain to be elucidated.

There may be two proton transfers in the carbonic anhydrase II-catalyzed mechanism of CO2 hydration that are important in catalysis, and both of these transfers are affected by the active-site zinc ion. The first (intramolecular) proton transfer may actually be a tautomerization between the intermediate and product forms of the bicarbonate anion (Fig. 28). This is believed to be a necessary step in the carbonic anhydrase II mechanism, due to a consideration of the reverse reaction. The cou-lombic attraction between bicarbonate and zinc is optimal when both oxygens of the delocalized anion face zinc, that is, when the bicarbonate anion is oriented with syn stereochemistry toward zinc (this is analogous to a syn-oriented carboxylate-zinc interaction see Fig. 28a). This energetically favorable interaction probably dominates the initial recognition of bicarbonate, but the tautomerization of zinc-bound bicarbonate is subsequently required for turnover in the reverse reaction (Fig. 28b). 

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