Radiation and chemical reactions that give rise to free radicals cause the formation of highly cytotoxic base propenals, degradation products DNA. Human glutathione transferases (GSTs; RX:glutathione R-transferase, EC 2.5.1.18) classes Alpha, Mu, Pi were shown promote conjugation with propenals related alkenes. GST P1-1 was particularly active in catalyzing propenal derivatives, adenine substrate giving highest activity. The catalytic efficiency (kcat/Km = 7.7 x 10(5) M-1.s-1) is so far...

The central role of protein kinases in signal transduction pathways has generated intense interest targeting these enzymes for a wide range therapeutic indications. Here we report method identifying and quantifying any biological sample or tissue from species. procedure relies on acyl phosphate-containing nucleotides, prepared biotin derivative ATP ADP. phosphate probes react selectively covalently at the binding sites least 75% known human kinases. Biotinylated peptide fragments labeled...

Nerve growth factor (NGF) is essential for the survival of both peripheral ganglion cells and central cholinergic neurons basal forebrain. The accelerated loss during Alzheimer's disease may be a determinant dementia in these patients therefore suggest therapeutic role NGF. However, NGF does not significantly penetrate blood-brain barrier, which makes its clinical utility dependent on invasive neurosurgical procedures. When conjugated to an antibody transferrin receptor, however, crossed...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTBleomycins: A Structural Model for Specificity, Binding, and Double Strand CleavageJoAnne Stubbe, John W. Kozarich, Wei Wu, Dana E. VanderwallView Author Information Departments of Chemistry Biology, Massachusetts Institute Technology, Cambridge, 02139, Merck Research Laboratories, P.O. Box 2000, Rahway, New Jersey 07065-0900Cite this: Acc. Chem. Res. 1996, 29, 7, 322–330Publication Date (Web):July 11, 1996Publication History Received1 January...

Bromovinyldeoxyuridine (BVdUrd) is a potent antiherpesvirus compound with low cytotoxicity. To gain an insight into its selectivity and mechanism of inhibition, we chemically synthesized the 5'-triphosphate BVdUrd, BVdUTP, tested effect on activities DNA polymerases [DNA nucleotidyltransferase (DNA directed), EC 2.7.7.7] two herpesviruses--i.e., herpes simplex virus type 1 (HSV-1) Epstein-Barr (EBV)--as well as cellular alpha, beta, gamma. The effects were determined under assay conditions...

Staphylococcus aureus H membranes were found to contain four major binding components: Mr = 115,000; 100,000 doublet; and 46,000. The low molecular weight protein bound penicillin reversibly was purified by prebinding with prior affinity chromatography. catalyzed transpeptidase carboxypeptidase reactions using di[14C]acetyl-L-lysyl-D-alanyl-D-alanine as the substrate glycine hydroxylamine acceptors. In addition, enzyme a penicillinase reaction. Kinetic analysis of these revealed similar Vmax...

Unlike other members of the MAPK family, ERK5 contains a large C-terminal domain with transcriptional activation capability in addition to an N-terminal canonical kinase domain. Genetic deletion is embryonic lethal, and tissue-restricted deletions have profound effects on erythroid development, cardiac function, neurogenesis. In addition, depletion antiinflammatory antitumorigenic. Small molecule inhibition has been shown promising activity cell animal models inflammation oncology. Here we...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTElectrophilic catalysis can explain the unexpected acidity of carbon acids in enzyme-catalyzed reactionsJohn A. Gerlt, John W. Kozarich, George L. Kenyon, and Paul G. GassmanCite this: J. Am. Chem. Soc. 1991, 113, 25, 9667–9669Publication Date (Print):December 1, 1991Publication History Published online1 May 2002Published inissue 1 December 1991https://pubs.acs.org/doi/10.1021/ja00025a039https://doi.org/10.1021/ja00025a039research-articleACS...

Finasteride is employed in treatment of benign prostatic hyperplasia man, where its target enzyme steroid 5α-reductase. It a novel, potent mechanism-based inhibitor the human prostate (type 2) isozyme. Although it accepted as an alternate substrate and ultimately reduced to dihydrofinasteride, this proceeds through enzyme-bound NADP−dihydrofinasteride adduct. processed with second-order rate constant, ki/Ki = 1 × 106 M-1 s-1, that approaches kcat/Km for reduction testosterone, 3 essentially...

Poly(dA*dU), which is specifically tritiated at the 1'-, 2'-(ribo configuration), 3'-, or 4'-position of deoxyuridine, has been synthesized and fate tritium determined upon degradation polymer by bleomycin, Fern), 02.No labilked from l'-3H-labeled as 'H20; however, resulting S-(uridin-l'-yl)-2-propenal (uracil propenal) expected specific activity.The 2'-3H-labeled affords 3H20 no label in uracil propenal.This result lack solvent incorporation into propenal suggest that proton abstraction...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTThe role of lysine 166 in the mechanism mandelate racemase from Pseudomonas putida: Mechanistic and crystallographic evidence for stereospecific alkylation by (R)-.alpha.-phenylglycidateJames A. Landro, John Gerlt, W. Kozarich, Carolyn Koo, Vibhakar J. Shah, George L. Kenyon, David Neidhart, Shigeo Fujita, Gregory PetskoCite this: Biochemistry 1994, 33, 3, 635–643Publication Date (Print):January 1, 1994Publication History Published online1 May...

Bleomycin is a metal- and oxygen-dependent DNA cleaver. The chemistry of damage has been proposed to involve rate-limiting abstraction the 4′-hydrogen. A fragment prepared that contains [4′- 2 H]thymidine residues high isotopic content. Primary kinetic isotope effects have directly observed at individual thymidine with sequencing technology.

Genome sequencing projects have uncovered many novel enzymes and enzyme classes for which knowledge of active site structure mechanism is limited. To facilitate mechanistic investigations the numerous encoded by prokaryotic eukaryotic genomes, new methods are needed to analyze function in samples high biocomplexity. Here, we describe a general strategy profiling sites whole proteomes that utilizes activity-based chemical probes coupled with gel-free analysis platform. We apply this identify...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTMechanism of bleomycin: evidence for a rate-determining 4'-hydrogen abstraction from poly(dA-dU) associated with the formation both free base and propenalJohn C. Wu, John W. Kozarich, JoAnne StubbeCite this: Biochemistry 1985, 24, 26, 7562–7568Publication Date (Print):December 17, 1985Publication History Published online1 May 2002Published inissue 17 December...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTIdentification and quantitation of the lesion accompanying base release in bleomycin-mediated DNA degradationLois E. Rabow, JoAnne Stubbe, John W. KozarichCite this: J. Am. Chem. Soc. 1990, 112, 8, 3196–3203Publication Date (Print):April 1, 1990Publication History Published online1 May 2002Published inissue 1 April 1990https://pubs.acs.org/doi/10.1021/ja00164a049https://doi.org/10.1021/ja00164a049research-articleACS PublicationsRequest reuse...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTPurification of the messenger RNA cap-binding protein using a new affinity mediumNancy R. Webb, Ravi V. J. Chari, Gia DePillis, John W. Kozarich, and Robert E. RhoadsCite this: Biochemistry 1984, 23, 2, 177–181Publication Date (Print):January 1, 1984Publication History Published online1 May 2002Published inissue 1 January 1984https://pubs.acs.org/doi/10.1021/bi00297a001https://doi.org/10.1021/bi00297a001research-articleACS PublicationsRequest reuse...

ADVERTISEMENT RETURN TO ISSUEPREVArticleSequence-Specific Double-Strand Cleavage of DNA by Fe-Bleomycin. 2. Mechanism and DynamicsM. J. Absalon, W. Wu, Kozarich, StubbeCite this: Biochemistry 1995, 34, 6, 2076–2086Publication Date (Print):February 14, 1995Publication History Published online1 May 2002Published inissue 14 February 1995https://pubs.acs.org/doi/10.1021/bi00006a030https://doi.org/10.1021/bi00006a030research-articleACS PublicationsRequest reuse permissionsArticle...

The solution structure of Co·Bleomycin (CoBLM) A2 green (the hydroperoxide form CoBLM) complexed with the self-complementary oligonucleotide d(CCAGGCCTGG) a cleavage site at C6 has been determined by 2D NMR spectroscopic methods and molecular dynamics calculations. Intermolecular NOEs (60 between CoBLM DNA) intramolecular (61 within green) have defined position orientation respect to its single binding in duplex. is stable analog activated BLM, Fe3+ (Sam, J. W.; Tang, X.-J.; Peisach, Am....

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTMandelate Racemase: Structure-Function Studies of a Pseudosymmetric EnzymeGeorge L. Kenyon, John A. Gerlt, Gregory Petsko, and W. KozarichCite this: Acc. Chem. Res. 1995, 28, 4, 178–186Publication Date (Print):April 1, 1995Publication History Published online1 May 2002Published inissue 1 April 1995https://pubs.acs.org/doi/10.1021/ar00052a003https://doi.org/10.1021/ar00052a003research-articleACS PublicationsRequest reuse permissionsArticle...

The structure of homogeneous Co·Bleomycin (CoBLM) A2 green (the hydroperoxide form CoBLM) has been determined using 2D NMR methods and molecular dynamics calculations. Previous studies Xu et al. (Xu, R. X.; Nettesheim, D.; Otvos, J. Petering, D. H. Biochemistry 1994, 33, 907−916) reported several possible structures for CoBLM compatible with their data acquired on a mixture brown forms. availability the pure green, which is stable months at neutral pH, allowed complete assignments 1H 13C...