Substrate mapping

Schematic diagram of the insulin receptor heterodimer in the activated state. IRS, insulin receptor substrate MAP, mitogen-activated protein P, phosphate tyr, tyrosine.

Tanaka, T. et al. 1985. Human leukocyte cathepsin G substrate mapping with 4-nitroanilides, chemical modification and effect of possible cofactors. Biochemistry, 24, 2040-2047. 

One key aspecf of applicafions of combinatorial libraries in chemical biology is fhe mapping of subsfrafe specificities for enzymes. Alfhough the early studies that use peptide libraries were focused on studying antibody epitopes (12), the substrate mapping for enzymes that transform peptides and the development of molecular probes found widespread interest. 

Active Site Model. Instead of developing an ideal substrate structure one also can try to picture the structure of the (unknown) active site of the enzyme by the method described above. Thus, substrates of varying size and polarity are used as probes to measure the dimensions of the active site. Therefore this approach has been denoted as substrate mapping [342, 343]. Such active site models are frequently employed and they usually resemble an arrangement of assumed sites or pockets which are usually box- or cave-shaped. A relatively reliable active-site model for PLE [344]... 

The experimental setup for the four-point probe techniques only requires an adequate probe head that satisfies the desired accuracy, which is mainly limited by the mechanical tolerances of the probe spacings, and a digital multimeter with a high input impedance of at least 10 Q. Today, several measurement systems are available on the market, due to their use in the semiconductor and display industries, ranging from a basically manual device to a fully automated system for substrate mapping. 

Scotts technology (17) uses fluid-bed (Wurster column) technology to apply polymeric coatings to a number of fertilizer substrates including urea, potassium nitrate, potassium sulfate, and monoammonium phosphate (MAP). The coating material is appHed as a water-borne latex onto the fluidized substrate. As the substrate is fluidized with warm air (40—50°C), water is driven off and the latex coalesces into a continuous film around the fertilizer particle. The particular latex compositions used have selected glass-transition and blocking temperatures, which enable quick removal of the water before the soluble fertilizer core dissolves. This obviates the need to use precoats prior to the latex appHcation. 

Spatial information about a system can be obtained by analyzing the spatial distribution of PL intensity. Fluorescent tracers may be used to image chemical uptake in biological systems. Luminescence profiles have proven useftil in the semiconductor industry for mapping impurity distributions, dislocadons, or structural homogeneity in substrate wafers or epilayers. Similar spatial infbrmadon over small regions is obtained by cathodoluminescence imaging. 

For a monolayer film, the stress-strain curve from Eqs. (103) and (106) is plotted in Fig. 15. For small shear strains (or stress) the stress-strain curve is linear (Hookean limit). At larger strains the stress-strain curve is increasingly nonlinear, eventually reaching a maximum stress at the yield point defined by = dT Id oLx x) = 0 or equivalently by c (q x4) = 0- The stress = where is the (experimentally accessible) static friction force [138]. By plotting T /Tlx versus o-x/o x shear-stress curves for various loads T x can be mapped onto a universal master curve irrespective of the number of strata [148]. Thus, for stresses (or strains) lower than those at the yield point the substrate sticks to the confined film while it can slip across the surface of the film otherwise so that the yield point separates the sticking from the slipping regime. By comparison with Eq. (106) it is also clear that at the yield point oo. 

With electron-deficient aromatic substrates (Entries 4 and 5), high yields and selectivities were observed, but enantioselectivities were variable and solvent-de-pendent (compare Entry 6 with 7 and see Section 1.2.1.3 for further discussion). With a,P-unsaturated tosylhydrazone salts, selectivities and yields were lower. The scope of this process has been extensively mapped out, enabling the optimum disconnection for epoxidation to be chosen [10]. 

MAPK cascades are composed of three cytoplasmic kinases, the MAPKKK, MAPKK, and MAPK, that are regulated by phosphorylation (Fig. 1) [1, 2]. The MAPKKK, also called MEKK for MEK kinase, is a serine/threonine kinase. Selective activation of MAPKKKs by upstream cellular stimuli results in the phosphorylation of MAPKK, also called MEK for MAP/ERK kinase by the MAPKKK. MAPKKK members are structurally diverse and are differentially regulated by specific upstream stimuli. The MAPKK is phosphorylated by the MAPKKK on two specific serine/ threonine residues in its activation loop. The MAPKK family members are dual specificity kinases capable of phosphorylating critical threonine and tyrosine residues in the activation loop of the MAPKs. MAPKKs have the fewest members in the MAPK signaling module. MAPKs are a family of serine/threonine kinases that upon activation by their respective MAPKKs, are capable of phosphorylating cytoplasmic substrates as well as... 

Propose a SECM experiment for mapping the distribution of an oxidase enzyme within a carbon composite surface. (Note that the enzyme generates hydrogen peroxide in the presence of its substrate and oxygen.)... 

Vmax and Km values using pga as a substrate at the optimum pH 4.1 were calculated as 500 U/mg and 0.15 mg/ml and 2000 U/mg and 0.15 mg/ml for PGI and PGII, respectively. Mode of action analysis revealed a random cleavage pattern for PGII while for PGI multiple attack on a single chain was observed. For PGII a partial subsite map was obtained. 

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