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Full-Text Articles in Physical Sciences and Mathematics
Structural Changes And Enhancements In Dnase I Footprinting Experiments?, Jerry Goodisman, James C. Dabrowiak
Structural Changes And Enhancements In Dnase I Footprinting Experiments?, Jerry Goodisman, James C. Dabrowiak
Chemistry - All Scholarship
In footprinting experiments, an increase in DNA cleavage with addition of ligand to a system may be due to a ligand-induced structural change. Ligand binding also enhances cleavage by displacing the cleavage agent from ligand-binding sites, thus increasing its concentration elsewhere. The theory and characteristics of this mass-action enhancement are given, and it is shown how it may be recognized. Results of DNase I footprinting of small oligomers, with actinomycin D as ligand, are analyzed to reveal which enhancements are due to mass action, and which can reasonably be ascribed to structural changes. Patterns in the footprinting plots from our …
Site-Specific Binding Constants For Actinomycin D On Dna Determined From Footprinting Studies, Jerry Goodisman, Robert Rehfuss, Brian Ward, James C. Dabrowiak
Site-Specific Binding Constants For Actinomycin D On Dna Determined From Footprinting Studies, Jerry Goodisman, Robert Rehfuss, Brian Ward, James C. Dabrowiak
Chemistry - All Scholarship
We report site-specific binding constants for the intercalating anticancer drug actinomycin D (Act-D), binding to a 139-base-pair restriction fragment from pBR 322 DNA. The binding constants are derived from analysis of footprinting experiments, in which the radiolabeled 139-mer is cleaved using DNase I, the cleavage products undergo gel electrophoresis, and, from the gel autoradiogram, spot intensities, proportional to amounts of cleaved fragments, are measured. A bound drug prevents DNase I from cleaving at -7 bonds, leading to decreased amounts of corresponding fragments. With the radiolabel on the 3’ end of the noncoding strand (A-label), we measured relative amounts of 54 …
Surface Tension Of A Charged And Polarized System, Jerry Goodisman
Surface Tension Of A Charged And Polarized System, Jerry Goodisman
Chemistry - All Scholarship
Usually, the formula for the surface tension of a planar charged and polarized interface is obtained from that for a system involving only short-range forces, y = - dz [p - px(z)] by replacing the tangential pressure p , by p , + E2/8u. Problems with this include (a) p, is no longer explicitly defined, (b) the electrostatic stress term E2/8 pi is not correct in general but only if polarization is proportional to density of polarizable species, (c) the derivation of the formula in terms of p and p, involves calculating the work to expand a volume containing the …
Formation Of Pentosidine During Nonenzymatic Browning Of Proteins By Glucose, Daniel G. Dyer, James A. Blackledge, Suzanne R. Thorpe, John W. Baynes
Formation Of Pentosidine During Nonenzymatic Browning Of Proteins By Glucose, Daniel G. Dyer, James A. Blackledge, Suzanne R. Thorpe, John W. Baynes
Faculty Publications
A fluorescent compound has been detected in proteins browned during Maillard reactions with glucose in vitro and shown to be identical to pentosidine, a pentose- derived fluorescent cross-link formed between arginine and lysine residues in collagen (Sell, D. R., and Monnier, V. M. (1989) J. Biol. Chem. 264, 21597- 2 1602). Pentosidine was the major fluorophore formed during nonenzymatic browning of ribonuclease and lysozyme by glucose, but accounted for <1% of nondisulfide cross-links in protein dimers formed during the reaction. Pentosidine was formed in greatest yields in reactions of pentoses with lysine and arginine in model systems but was also formed from glucose, fructose, ascorbate, Amadori compounds, 3-deoxyglucosone, and other sugars. Pentosidine was not formed from peroxidized polyunsaturated fatty acids or malondialdehyde. Its formation from carbohydrates was inhibited under nitrogen or anaerobic conditions and by aminoguanidine, an inhibitor of advanced glycation and browning reactions. Pentosidine was detected in human lens proteins, where its concentration increased gradually with age, but it did not exceed trace concentrations (55 Fmol/mol lysine), even in the 80-year-old lens. Although its precise carbohydrate source in vivo is uncertain and it is present in only trace concentrations in tissue proteins, pentosidine appears to be a useful biomarker for assessing cumulative damage to proteins by nonenzymatic browning reactions with carbohydrates.
Decrease In Skin Collagen Glycation With Improved Glycemic Control In Patients With Insulin-Dependent Diabetes Mellitus, Timothy J. Lyons, Karen E. Bailie, Daniel G. Dyer, John A. Dunn, John W. Baynes
Decrease In Skin Collagen Glycation With Improved Glycemic Control In Patients With Insulin-Dependent Diabetes Mellitus, Timothy J. Lyons, Karen E. Bailie, Daniel G. Dyer, John A. Dunn, John W. Baynes
Faculty Publications
Glycation, oxidation, and nonenzymatic browning of protein have all been implicated in the development of diabetic complications. The initial product of glycation of protein, fructoselysine (FL), undergoes further reactions, yielding a complex mixture of browning products, including the fluorescent lysine-arginine cross-link, pentosidine. Alternatively, FL may be cleaved oxidatively to form N(epsilon)-(carboxymethyl)lysine (CML), while glycated hydroxylysine, an amino-acid unique to collagen, may yield N(epsilon)-(carboxymethyl)hydroxylysine (CMhL). We have measured FL, pentosidine, fluorescence (excitation = 328 nm, emission = 378 nm), CML, and CMhL in insoluble skin collagen from 14 insulin-dependent diabetic patients before and after a 4-mo period of intensive therapy to …