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352. Microfluidic Chip–Based Online Screening Coupled to Mass Spectrometry.

Author

Iyer, Janaki Krishnamoorthy, Otvos, Reka A., Kool, Jeroen, and Kini, R. Manjunatha

Abstract

Thrombin and factor Xa (FXa) are critical enzymes of the blood coagulation cascade and are excellent targets of anticoagulant agents. Natural sources present an array of anticoagulants that can be developed as antithrombotic drugs. High-resolution, online screening techniques have been developed for the identification of drug leads from complex mixtures. In this study, we have developed and optimized a microfluidic online screening technique coupled to nano–liquid chromatography (LC) and in parallel with a mass spectrometer for the identification of thrombin and FXa inhibitors in mixtures. Inhibitors eluting from the nano-LC were split postcolumn in a 1:1 ratio; half was fed into a mass spectrometer (where its mass is detected), and the other half was fed into a microfluidic chip (which acts as a microreactor for the online assays). With our platform, thrombin and FXa inhibitors were detected in the assay in parallel with their mass identification. These methods are suitable for the identification of inhibitors from sample amounts as low as sub-microliter volumes. [ABSTRACT FROM AUTHOR]

Published
2016
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360. A Defined α-Helix in the Bifunctional O-Glycosylated Natriuretic Peptide TcNPa from the Venom of Tropidechis carinatus.

Author

Reeks, Timothy, Jones, Alun, Brust, Andreas, Sridharan, Sindhuja, Corcilius, Leo, Wilkinson, Brendan L., Thaysen ‐ Andersen, Morten, Payne, Richard J., Kini, R. Manjunatha, Daly, Norelle L., and Alewood, Paul F.

Subjects
NATRIURETIC peptides, SNAKE venom, PEPTIDE synthesis, GLYCOSIDASES, GUANYLATE cyclase, GLYCOSYLATION, PHENYLALANINE, HEART physiology
Abstract

Natriuretic peptides (NP) play important roles in human cardiac physiology through their guanylyl cyclase receptors NPR-A and NPR-B. Described herein is a bifunctional O-glycosylated natriuretic peptide, TcNPa, from Tropidechis carinatus venom and it unusually targets both NPR-A and NPR-B. Characterization using specific glycosidases and ETD-MS identified the glycan as galactosyl-β(1-3)- N-acetylgalactosamine (Gal-GalNAc) and was α-linked to the C-terminal threonine residue. TcNPa contains the characteristic NP 17-membered disulfide ring with conserved phenylalanine and arginine residues. Both glycosylated and nonglycosylated forms were synthesized by Fmoc solid-phase peptide synthesis and NMR analysis identified an α-helix within the disulfide ring containing the putative pharmacophore for NPR-A. Surprisingly, both forms activated NPR-A and NPR-B and were relatively resistant towards proteolytic degradation in plasma. This work will underpin the future development of bifunctional NP peptide mimetics. [ABSTRACT FROM AUTHOR]

Published
2015
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365. Denmotoxin, a Three-finger Toxin from the Colubrid Snake Boiga dendrophila (Mangrove Catsnake) with Bird-specific Activity

Author

Pawlak, Joanna, primary, Mackessy, Stephen P., additional, Fry, Bryan G., additional, Bhatia, Madhav, additional, Mourier, Gilles, additional, Fruchart-Gaillard, Carole, additional, Servent, Denis, additional, Ménez, Renée, additional, Stura, Enrico, additional, Ménez, André, additional, and Kini, R. Manjunatha, additional

Published
2006
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392. Preface

Author

Kini, R. Manjunatha, primary, Marsh, Neville A., additional, Markland, Francis S., additional, and Broady, Kevin W., additional

Published
2005
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