Enzyme facilitated diffusion

FIGURE 10.3 Passive diffusion and facilitated diffusion may be distinguished graphically. The plots for facilitated diffusion are similar to plots of enzyme-catalyzed processes (Chapter 14) and they display saturation behav-... 

Mutations in bacteria and mammalian cells (including some that result in human disease) have supported these conclusions. Facilitated diffusion and active transport resemble a substrate-enzyme reaction except that no covalent interaction occurs. These points of resemblance are as follows (1) There is a specific binding site for the solute. (2) The carrier is saturable, so it has a maximum rate of transport (V Figure 41-11). (3) There is a binding constant (Al) ) for the solute, and... 

Conditional stability constants have been determined for cadmium binding to humic acid in freshwater, log Kk 6.5 [27], which may be comparable to binding to humic acid coated particles. The experiments demonstrated the importance of cadmium uptake from particles rather than from the dissolved phase. The authors recognised that the overall conclusion was similar to previous studies [28], but there remain inconsistencies in the uptake levels which may be related to the heterogeneity of the systems. Uptake from the intestine into the mucosal cells was not investigated. It was presumed that the material was digested extracellularly by hydrolytic enzymes and the released metal was taken up by facilitated diffusion. 

Facilitated diffusion within organisms takes place when carriers or proteins residing within membranes—ion channels, for instance—organize the movement of ions from one location to another. This diffusion type is a kinetic, not thermodynamic, effect in which a for the transfer is lowered and the rate of diffusion is accelerated. Facilitated diffusion channels organize ion movements in both directions, and the process can be inhibited both competitively and noncompetitively. It is known that most cells maintain open channels for K+ most of the time and closed channels for other ions. Potassium-ion-dependent enzymes include NaVK+ ATPases (to be discussed in Section 5.4.1), pyruvate kinases, and dioldehydratases (not to be discussed further). 

Carrier-mediated passage of a molecular entity across a membrane (or other barrier). Facilitated transport follows saturation kinetics ie, the rate of transport at elevated concentrations of the transportable substrate reaches a maximum that reflects the concentration of carriers/transporters. In this respect, the kinetics resemble the Michaelis-Menten behavior of enzyme-catalyzed reactions. Facilitated diffusion systems are often stereo-specific, and they are subject to competitive inhibition. Facilitated transport systems are also distinguished from active transport systems which work against a concentration barrier and require a source of free energy. Simple diffusion often occurs in parallel to facilitated diffusion, and one must correct facilitated transport for the basal rate. This is usually evident when a plot of transport rate versus substrate concentration reaches a limiting nonzero rate at saturating substrate While the term passive transport has been used synonymously with facilitated transport, others have suggested that this term may be confused with or mistaken for simple diffusion. See Membrane Transport Kinetics... 

There are, however, various types of active transport systems, involving protein carriers and known as uniports, symports, and antiports as indicated in Figure 3.7. Thus, symports and antiports involve the transport of two different molecules in either the same or a different direction. Uniports are carrier proteins, which actively or passively (see section "Facilitated Diffusion") transport one molecule through the membrane. Active transport requires a source of energy, usually ATP, which is hydrolyzed by the carrier protein, or the cotransport of ions such as Na+ or H+ down their electrochemical gradients. The transport proteins usually seem to traverse the lipid bilayer and appear to function like membrane-bound enzymes. Thus, the protein carrier has a specific binding site for the solute or solutes to be transferred. For example, with the Na+/K+ ATPase antiport, the solute (Na+) binds to the carrier on one side of... 

Membrane proteins lower the activation energy for transport of polar compounds and ions by providing an alternative path through the bilayer for specific solutes. Proteins that bring about this facilitated diffusion, or passive transport, are not enzymes in the usual sense their substrates are moved from one compartment to another, but are not chemically altered. Membrane proteins that speed the movement of a solute across a membrane by facilitating diffusion are called transporters or permeases. 

The amino acid sequences of numerous transport proteins have been elucidated during the last 10 years. As with conventional enzymes, molecular biological techniques now make it possible to modify individual amino acid residues in the proteins and thus to probe the roles these residues play in catalysis of facilitated diffusion or active transport. 

Biological membranes present a barrier to the free transport of cations, as the hydrophilic, hydrated cations cannot cross the central hydrophobic region of the membrane which is formed by the hydrocarbon tails of the lipids in the bilayer. Specific mechanisms thus have to be provided for the transport of cations, which therefore allow for the introduction of controls. Such translocation processes may involve the active transport of cations against the concentration gradient with expenditure of energy via the hydrolysis of ATP. These ion pumps involve enzyme activity. Alternatively, facilitated diffusion may occur in which the cation is assisted to cross the hydrophobic barrier. Such diffusion will follow the concentration gradient until concentrations either side of... 

Facilitated diffusion has certain general characteristics. As already mentioned, the net flux is toward a lower chemical potential. (According to the usual definition, active transport is in the energetically uphill direction active transport may use the same carriers as those used for facilitated diffusion.) Facilitated diffusion causes fluxes to be larger than those expected for ordinary diffusion. Furthermore, the transporters can exhibit selectivity (Fig. 3-17) that is, they can be specific for certain molecules solute and not bind closely related ones, similar to the properties of enzymes. In addition, carriers in facilitated diffusion become saturated when the external concentration of the solute transported is raised sufficiently, a behavior consistent with Equation 3.28. Finally, because carriers can exhibit competition, the flux density of a solute entering a cell by facilitated diffusion can be reduced when structurally similar molecules are added to the external solution. Such molecules compete for the same sites on the carriers and thereby reduce the binding and the subsequent transfer of the original solute into the cell. 

Dietary biotin bound to avidin (Section 11.6) is unavailable, but intravenously administered avidin-biotin is biologically active. Cells in culture are not inhibited by the addibon of avidin to the culmre medium, and can take up the avidin-biotin complex by pinocytosis followed by lysosomal hydrolysis, releasing free biotin. Unlike other B vitamins, for which concentrative uptake into tissues is achieved by facilitated diffusion, followed by metabolic trapping, the incorporation of biotin into enzymes is slow and cannot be considered part of the uptake process. 

Dietary NAD and NADP are hydrolyzed by enzymes, such as NAD glycohydrolase, in the intestinal mucosa to release nicotinamide, which together with any nicotinic acid is rapidly absorbed in both the stomach and intestine by an Na -dependent facilitated diffusion at low concentrations and passive diffusion at higher concentrations. Nicotinamide is the main circulating form in the plasma, either postabsorption or by release from hydrolyzed liver NAD, and this can be taken up by most tissue requiring NAD by simple diffusion, though there is evidence of a facilitated transport in the erythrocyte. ... 

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