ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTKinetic mechanism of DNA polymerase I (Klenow)R. D. Kuchta, V. Mizrahi, P. A. Benkovic, K. Johnson, and S. J. BenkovicCite this: Biochemistry 1987, 26, 25, 8410–8417Publication Date (Print):December 1, 1987Publication History Published online1 May 2002Published inissue 1 December 1987https://pubs.acs.org/doi/10.1021/bi00399a057https://doi.org/10.1021/bi00399a057research-articleACS PublicationsRequest reuse permissionsArticle...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTKinetic mechanism whereby DNA polymerase I (Klenow) replicates with high fidelityRobert D. Kuchta, Patricia Benkovic, and Stephen J. BenkovicCite this: Biochemistry 1988, 27, 18, 6716–6725Publication Date (Print):September 6, 1988Publication History Published online1 May 2002Published inissue 6 September 1988https://pubs.acs.org/doi/10.1021/bi00418a012https://doi.org/10.1021/bi00418a012research-articleACS PublicationsRequest reuse...

ABSTRACT Global surveillance of influenza is critical for improvements in disease management and especially important early detection, rapid intervention, a possible reduction the impact an pandemic. Enhanced requires rapid, robust, inexpensive analytical techniques capable providing detailed analysis virus strains. Low-density oligonucleotide microarrays with highly multiplexed “signatures” viruses offer many desired characteristics. However, high mutability represents design challenge. In...

Proteolysis targeting chimeras (PROTACs) hijacking the cereblon (CRBN) E3 ubiquitin ligase have emerged as a novel paradigm in drug development. Herein we found that linker attachment points of CRBN ligands highly affect their aqueous stability and neosubstrate degradation features. This work provides blueprint for assembly future heterodimeric CRBN-based degraders with tailored properties.

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTAcyclic Guanosine Analogs Inhibit DNA Polymerases .alpha., .delta., and .epsilon. with Very Different Potencies Have Unique Mechanisms of ActionDiane D. Ilsley, Suk-Hee Lee, Wayne H. Miller, Robert KuchtaCite this: Biochemistry 1995, 34, 8, 2504–2510Publication Date (Print):February 28, 1995Publication History Published online1 May 2002Published inissue 28 February...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTDNA polymerase .epsilon.: Aphidicolin inhibition and the relationship between exonuclease activityChung Hui Cheng Robert D. KuchtaCite this: Biochemistry 1993, 32, 33, 8568–8574Publication Date (Print):August 24, 1993Publication History Published online1 May 2002Published inissue 24 August 1993https://pubs.acs.org/doi/10.1021/bi00084a025https://doi.org/10.1021/bi00084a025research-articleACS PublicationsRequest reuse permissionsArticle...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTMechanism of DNA polymerase .alpha. inhibition by aphidicolinRobert Sheaff, Diane Ilsley, and Robert KuchtaCite this: Biochemistry 1991, 30, 35, 8590–8597Publication Date (Print):September 3, 1991Publication History Published online1 May 2002Published inissue 3 September 1991https://pubs.acs.org/doi/10.1021/bi00099a014https://doi.org/10.1021/bi00099a014research-articleACS PublicationsRequest reuse permissionsArticle...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTMechanism of calf thymus DNA primase: Slow initiation, rapid polymerization, and intelligent terminationRobert J. Sheaff Robert D. KuchtaCite this: Biochemistry 1993, 32, 12, 3027–3037Publication Date (Print):March 1, 1993Publication History Published online1 May 2002Published inissue 1 March 1993https://pubs.acs.org/doi/10.1021/bi00063a014https://doi.org/10.1021/bi00063a014research-articleACS PublicationsRequest reuse permissionsArticle...

We have obtained a soluble system that carries out this conversion as well the hydration of acrylate to lactate and reduction propionate.3-Pentynyl-CoA inhibits propionate, but not by cell extracts.The probably involves CoA esters.When [/3-2Hs] is used substrate, rate propionate formation reduced 1.8-foldy methyl group resulting has lost 1.4 deuterium atoms.These results are consistent with intermediate (acrylyl-CoA) in wlactate propionate.Two proteins, which we designate E I 11, were...

Protein ubiquitination plays an important role in the regulation of almost every aspect eukaryotic cellular function; therefore, its destabilization is often observed most human diseases and cancers. Consequently, developing inhibitors system for treatment cancer has been a recent area interest. Currently, only few classes compounds have discovered to inhibit ubiquitin-activating enzyme (E1) one class relatively selective E1 inhibition cells. We now report that Largazole ester ketone analogs...

The p58 subunit of human DNA primase contains a region, M288−K344, that is homologous to part the 8 kDa domain polymerase β. Since regions protein are highly conserved evolutionarily often play important catalytic functions, we examined effects mutating this region on activity. Deleting M288−L313 results in binds p49 but cannot support primer synthesis any template when assays only contain Mg2+ as divalent metal. Including Mn2+, metal stimulates initiation synthesis, now allows enzyme...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTMechanism of DNA replication fidelity for three mutants polymerase I: Klenow fragment KF(exo+), KF(polA5), and KF(exo-)Bryan T. Eger, Robert D. Kuchta, Steven S. Carroll, Patricia A. Benkovic, Michael E. Dahlberg, Catherine M. Joyce, Stephen J. BenkovicCite this: Biochemistry 1991, 30, 5, 1441–1448Publication Date (Print):February 1991Publication History Published online1 May 2002Published inissue 5 February...

The performance of a diagnostic microarray (the MChip assay) for influenza was compared in blind study to that viral culture, reverse transcription (RT)-PCR, and the QuickVue Influenza A+B test. patient sample data set composed 102 respiratory secretion specimens collected between 29 December 2005 2 February 2006 at Scott & White Hospital Clinic Temple, Texas. Samples were from wide range age groups by using direct collection, nasal/nasopharyngeal swabs, or nasopharyngeal aspiration. Viral...

Calf thymus DNA primase was examined to determine the kinetic parameters that define its unusual processivity. At 37 degrees C, major products were 8-9 and 2-3 nucleotides long. The 2-mer predominant product when considered on a molar basis. each polymerization cycle en route synthesis of unit length primer (7-10 nucleotides), processivity defined by competition enzyme dissociation with ATP binding as well an independent step(s). Reducing temperature 25 C had relatively little effect...

The design and characterization of a low-density microarray for subtyping influenza A is presented. consisted 15 distinct oligonucleotides designed to target only the matrix gene segment A. An artificial neural network was utilized automate image interpretation. trained recognize fluorescence patterns 68 known viruses subsequently used identify 53 unknowns in blind study that included 39 human patient samples 14 negative control samples. assay exhibited clinical sensitivity 95% specificity 92%.

Extending earlier studies on iodine catalysis, experimental investigations show that various halogen-bond donors can also be employed to accelerate the Michael addition between trans-crotonophenone and indole. Solvent as well counteranion effects have been analyzed, kinetic computational provide additional insights into mode of activation.

Fluorescent RNA is an important analytical tool in medical diagnostics, cytochemistry, and aptamer development. We have synthesized the fluorescent ribonucleotide analogue 1,3-diaza-2-oxophenothiazine-ribose-5'-triphosphate (tCTP) tested it as substrate for T7 polymerase transcription reactions, a convenient route generating vitro. When transcribing guanine, incorporates tCTP with 2-fold higher catalytic efficiency than CTP efficiently polymerizes additional NTPs onto tC. Remarkably, does...

Protein degradation via the ubiquitin-proteasome pathway is important for a diverse number of cellular processes ranging from cell signaling to development. Disruption ubiquitin occurs in variety human diseases, including several cancers and neurological disorders. Excessive proteolysis tumor suppressor proteins, such as p27, numerous aggressive tumors. To discover small-molecule inhibitors that potentially prevent p27 degradation, we developed series screening assays, cell-based screen...

Recent in vitro work with Golgi-enriched membranes showed that 3′-azidothymidine-5′-monophosphate (AZTMP), the primary intracellular metabolite of 3′-azidothymidine (AZT), is a potent inhibitor glycosylation reactions (Hall et al.(1994) J. Biol. Chem. 269, 14355-14358) and predicted AZT treatment whole cells should cause similar inhibition. In this report, we verify prediction by showing K562 inhibits lipid protein glycosylation. dramatically alters pattern glycosphingolipid biosynthesis,...