Abstract
Proteomics analysis was a powerful technology for characterizing proteins and protein posttranslational modification (PTMs). Recently, many anther and pollen-related proteomic analyses have been reported, which have expanded our understanding of anther and pollen development and regulation. In this chapter, we describe a detailed, optimized protocol for the separation, digestion, tagging, and subsequent mass spectrometry-based identification and quantification of proteins and phosphoproteins from anther and pollen.
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References
Ma H (2005) Molecular genetic analyses of microsporogenesis and microgametogenesis in flowering plants. Annu Rev Plant Biol 56:393–434
Chang F, Wang Y, Wang S, Ma H (2011) Molecular control of microsporogenesis in Arabidopsis. Curr Opin Plant Biol 14:66–73
Huang MD, Hsing YI, Huang AH (2011) Transcriptomes of the anther sporophyte: availability and uses. Plant Cell Physiol 52:1459–1466
Sanders PM, Bui AQ, Weterings K, Mcintire KN, Hsu YC, Pei YL et al (1999) Anther developmental defects in Arabidopsis thaliana male-sterile mutants. Sex Plant Reprod 11:297–322
Wilkins MR, Sanchez JC, Gooley AA, Appel RD, Humphery-Smith I, Hochstrasser DF et al (1996) Progress with proteome projects: why all proteins expressed by a genome should be identified and how to do it. Biotechnol Genet Eng Rev 13:19–50
Pandey A, Mann M (2000) Proteomics to study genes and genomes. Nature 405:837–846
Gevaert K, Van Damme J, Goethals M, Thomas GR, Hoorelbeke B, Demol H et al (2002) Chromatographic isolation of methionine-containing peptides for gel-free proteome analysis: identification of more than 800 Escherichia coli proteins. Mol Cell Proteomics 1:896–903
Roe MR, Griffin TJ (2006) Gel-free mass spectrometry-based high throughput proteomics: tools for studying biological response of proteins and proteomes. Proteomics 6:4678–4687
Van den Bergh G, Arckens L (2005) Recent advances in 2D electrophoresis: an array of possibilities. Expert Rev Proteomics 2:243–252
Vercauteren FG, Bergeron JJ, Vandesande F, Arckens L, Quirion R (2004) Proteomic approaches in brain research and neuropharmacology. Eur J Pharmacol 500:385–398
Grobei MA, Qeli E, Brunner E, Rehrauer H, Zhang R, Roschitzki B, Basler K, Ahrens CH, Grossniklaus U (2009) Deterministic protein inference for shotgun proteomics data provides new insights into Arabidopsis pollen development and function. Genome Res 19:1786–1800
Mayank P, Grossman J, Wuest S, Boisson-Dernier A, Roschitzki B, Nanni P, Nuhse T, Grossniklaus U (2012) Characterization of the phosphoproteome of mature Arabidopsis pollen. Plant J 72:89–101
Ye J, Zhang Z, Long H, Zhang Z, Hong Y, Zhang X et al (2015) Proteomic and phosphoproteomic analyses reveal extensive phosphorylation of regulatory proteins in developing rice anthers. Plant J 84:527–544
Ye J, Zhang Z, You C, Zhang X, Lu J, Ma H (2016) Abundant protein phosphorylation potentially regulates Arabidopsis anther development. J Exp Bot 67:4993–5008
Zhang Z, Hu M, Feng X, Gong A, Cheng L, Yuan H (2017) Proteomes and phosphoproteomes of anther and pollen: availability and progress. Proteomics 17(20). https://doi.org/10.1002/pmic.201600458
Wijeratne AJ, Zhang W, Sun Y, Liu W, Albert R, Zheng Z et al (2007) Differential gene expression in Arabidopsis wild-type and mutant anthers: insights into anther cell differentiation and regulatory networks. Plant J 52:14–29
Zhang D, Luo X, Zhu L (2011) Cytological analysis and genetic control of rice anther development. J Genet Genomics 38:379–390
Larsen MR, Thingholm TE, Jensen ON, Roepstorff P, Jorgensen TJ (2005) Highly selective enrichment of phosphorylated peptides from peptide mixtures using titanium dioxide microcolumns. Mol Cell Proteomics 4:873–886
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Zhang, C., Feng, X., Hu, M., Zhang, Z. (2020). How to Study the Proteomes and Phosphoproteomes of Anther and Pollen. In: Pradillo, M., Heckmann, S. (eds) Plant Meiosis. Methods in Molecular Biology, vol 2061. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9818-0_18
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DOI: https://doi.org/10.1007/978-1-4939-9818-0_18
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