Mass Spectrometry for Proteomics-Based Investigation Using the Zebrafish Vertebrate Model System

The zebrafish (Danio rerio ) is now the pre-eminent vertebrate model system for clarification of the roles of specific genes and signaling pathways in development. The zebrafish genome will be completely sequenced within the next 1—2 years. Together with the substantial historical database regarding basic developmental biology, toxicology, and gene transfer, the rich foundation of molecular genetic and genomic data makes zebrafish a powerful model system for clarifying mechanisms in toxicity. In contrast to the highly advanced knowledge base on molecular developmental genetics in zebrafish, our database regarding infectious and noninfectious diseases and pathologic lesions in zebrafish lags far behind the information available on most other domestic mammalian and avian species, particularly rodents. Currently, minimal data are available regarding spontaneous neoplasm rates or spontaneous aging lesions in any of the commonly used wild-type or mutant lines of zebrafish. Therefore, to fully utilize the potential of zebrafish as an animal model for understanding human development, disease, and toxicology we must greatly advance our knowledge on zebrafish diseases and pathology.

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A Fundamental Study of Amadori Rearrangement Products in Reducing Sugar-Amino Acid Model System by Electrospray Ionization Mass Spectrometry and Computation

Asian Journal of Chemistry ◽

10.14233/ajchem.2014.16078 ◽

2014 ◽

Vol 26 (10) ◽

pp. 2941-2944

Author(s):

Y.F. Zhang ◽

E.D. Ruan ◽

H. Wang ◽

Y.Y. Ruan ◽

G.J. Shao ◽

...

Keyword(s):

Mass Spectrometry ◽

Amino Acid ◽

Electrospray Ionization ◽

Electrospray Ionization Mass Spectrometry ◽

Model System ◽

Reducing Sugar ◽

Ionization Mass Spectrometry ◽

Ionization Mass ◽

Fundamental Study ◽

Amadori Rearrangement

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Transferrin as a model system for method development to study structure, dynamics and interactions of metalloproteins using mass spectrometry

Biochimica et Biophysica Acta (BBA) - General Subjects ◽

10.1016/j.bbagen.2011.06.019 ◽

2012 ◽

Vol 1820 (3) ◽

pp. 417-426 ◽

Cited By ~ 20

Author(s):

Igor A. Kaltashov ◽

Cedric E. Bobst ◽

Mingxuan Zhang ◽

Rachael Leverence ◽

Dmitry R. Gumerov

Keyword(s):

Mass Spectrometry ◽

Method Development ◽

Model System ◽

Structure Dynamics

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Demonstrating Enhanced Throughput of RapidFire Mass Spectrometry through Multiplexing Using the JmjD2d Demethylase as a Model System

CrossRef Listing of Deleted DOIs ◽

10.1177/1087057113496276 ◽

2013 ◽

Vol 19 (2) ◽

pp. 278-286 ◽

Cited By ~ 27

Author(s):

Melanie Leveridge ◽

Rachel Buxton ◽

Argyrides Argyrou ◽

Peter Francis ◽

Bill Leavens ◽

...

Keyword(s):

Mass Spectrometry ◽

Enzymatic Activity ◽

High Throughput ◽

Fold Increase ◽

Model System ◽

Analysis Time ◽

Sample Handling ◽

High Throughput Screens ◽

Pooled Samples ◽

Large Compound

Using mass spectrometry to detect enzymatic activity offers several advantages over fluorescence-based methods. Automation of sample handling and analysis using platforms such as the RapidFire (Agilent Technologies, Lexington, MA) has made these assays amenable to medium-throughput screening (of the order of 100,000 wells). However, true high-throughput screens (HTS) of large compound collections (>1 million) are still considered too time-consuming to be feasible. Here we propose a simple multiplexing strategy that can be used to increase the throughput of RapidFire, making it viable for HTS. The method relies on the ability to analyze pooled samples from several reactions simultaneously and to deconvolute their origin using “mass-tagged” substrates. Using the JmjD2d H3K9me3 demethylase as a model system, we demonstrate the practicality of this method to achieve a 4-fold increase in throughput. This was achieved without any loss of assay quality. This multiplex strategy could easily be scaled to give even greater reductions in analysis time.

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