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Absolute Quantification of Proteins in the Eye of Drosophila melanogaster.

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Raghuraman,  Bharath Kumar
Max Planck Institute for Molecular Cell Biology and Genetics, Max Planck Society;

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Hebbar,  Sarita
Max Planck Institute for Molecular Cell Biology and Genetics, Max Planck Society;

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Kumar,  Mukesh
Max Planck Institute for Molecular Cell Biology and Genetics, Max Planck Society;

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Moon,  HongKee
Max Planck Institute for Molecular Cell Biology and Genetics, Max Planck Society;

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Henry,  Ian
Max Planck Institute for Molecular Cell Biology and Genetics, Max Planck Society;

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Knust,  Elisabeth
Max Planck Institute for Molecular Cell Biology and Genetics, Max Planck Society;

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Shevchenko,  Andrej
Max Planck Institute for Molecular Cell Biology and Genetics, Max Planck Society;

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Citation

Raghuraman, B. K., Hebbar, S., Kumar, M., Moon, H., Henry, I., Knust, E., et al. (2020). Absolute Quantification of Proteins in the Eye of Drosophila melanogaster. Proteomics, 20(23): e1900049. doi:10.1002/pmic.201900049.


Cite as: https://hdl.handle.net/21.11116/0000-0008-A254-2
Abstract
Absolute (molar) quantification of proteins determines their molar ratios in complexes, networks and metabolic pathways. We employed MS Western workflow to determine molar abundances of proteins potentially critical for morphogenesis and phototransduction (PT) in eyes of Drosophila melanogaster. We used a single chimeric 264 kDa protein standard that covers, in total, 197 peptides from 43 proteins. The majority of proteins were independently quantified with 2 to 4 proteotypic peptides with the coefficient of variation of less than 15%, better than 1000-fold dynamic range and sub-femtomole sensitivity. We determined molar abundances of the components of the PT machinery and the rhabdomere, the photosensitive organelle of the fly eye, and how they changed when rhabdomere morphogenesis is perturbed by genetic manipulation of the evolutionary conserved gene crumbs (crb). Data are available via ProteomeXchange with identifier PXD018001 This article is protected by copyright. All rights reserved.