Kinetic Characterization of Prenyl-Flavin Synthase from Saccharomyces cerevisiae

December 27th, 2017 by Nattapol Arunrattanamook and E. Neil G. Marsh

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Biochemistry
DOI: 10.1021/acs.biochem.7b01131

Seven Year Itch: Pan-Assay Interference Compounds (PAINS) in 2017Utility and Limitations

December 26th, 2017 by Jonathan B. Baell and J. Willem M. Nissink

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ACS Chemical Biology
DOI: 10.1021/acschembio.7b00903

The CDC50A extracellular domain is required for forming a functional complex with and chaperoning phospholipid flippases to the plasma membrane [Membrane Biology]

December 24th, 2017 by Katumori Segawa, Sachiko Kurata, Shigekazu Nagata

Flippases are enzymes that translocate phosphatidylserine (PtdSer) and phosphatidyl- ethanolamine (PtdEtn) from the outer to the inner leaflet in the lipid bilayer of the plasma membrane, leading to the asymmetric distribution of aminophospholipids in the membrane. One mammalian phospholipid flippase at the plasma membrane is ATP11C, a type IV P-type ATPase (P4-ATPase) that forms a hetero-complex with the transmembrane protein CDC50A. However, the structural features in CDC50A that support the function of ATP11C and other P4-ATPases have not been characterized. Here, using error-prone PCR-mediated mutagenesis of human CDC50A cDNA followed by functional screening and deep sequencing, we identified 14 amino acid residues that affect ATP11C’s flippase activity. These residues were all located in CDC50A’s extracellular domain and were evolutionarily well conserved. Most of the mutations decreased CDC50A’s ability to chaperone ATP11C and other P4-ATPases to their destinations. The CDC50A mutants failed to form a stable complex with ATP11C and could not induce ATP11C’s PtdSer-dependent ATPase activity. Notably, one mutant variant could form a stable complex with ATP11C and transfer ATP11C to the plasma membrane, yet the ATP11C complexed with this CDC50A variant had very weak or little PtdSer- or PtdEtn-dependent ATPase activity. These results indicated that the extracellular domain of CDC50A has important roles both in CDC50A’s ability to chaperone ATP11C to the plasma membrane and in inducing ATP11C’s ATP hydrolysis–coupled flippase activity.
  • Posted in Journal of Biological Chemistry, Publications
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Complexity in pH-Dependent Ribozyme Kinetics: Dark pKa Shifts and Wavy Rate–pH Profiles

December 22nd, 2017 by Erica A. Frankel and Philip C. Bevilacqua

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Biochemistry
DOI: 10.1021/acs.biochem.7b00784

RNA Thermometers for the PURExpress System

December 22nd, 2017 by Fredrik W. Sadler, Igor Dodevski and Casim A. Sarkar

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ACS Synthetic Biology
DOI: 10.1021/acssynbio.7b00294

Development of a Pigment-Based Whole-Cell Zinc Biosensor for Human Serum

December 22nd, 2017 by Daniel M. Watstein and Mark P. Styczynski

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ACS Synthetic Biology
DOI: 10.1021/acssynbio.7b00292

A Genetic Toolbox for Modulating the Expression of Heterologous Genes in the Cyanobacterium Synechocystis sp. PCC 6803

December 22nd, 2017 by Bo Wang, Carrie Eckert, Pin-Ching Maness and Jianping Yu

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ACS Synthetic Biology
DOI: 10.1021/acssynbio.7b00297
  • Posted in ACS Synthetic Biology, Publications
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Where Informatics Lags Chemistry Leads

December 22nd, 2017 by Rahul Kaushik, Ankita Singh and B. Jayaram

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Biochemistry
DOI: 10.1021/acs.biochem.7b01073

Structural Features and Domain Movements Controlling Substrate Binding and Cofactor Specificity in Class II HMG-CoA Reductase

December 21st, 2017 by Bradley R. Miller and Yan Kung

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Biochemistry
DOI: 10.1021/acs.biochem.7b00999

Expression system for structural and functional studies of human glycosylation enzymes

December 18th, 2017 by Kelley W Moremen

Expression system for structural and functional studies of human glycosylation enzymes

Expression system for structural and functional studies of human glycosylation enzymes, Published online: 18 December 2017; doi:10.1038/nchembio.2539

A modular protein expression system enables the structural and functional characterization of human glycosyltransferases, glycoside hydrolases and other carbohydrate-modifying enzymes.
  • Posted in Nat Chem Biol, Publications
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