Efficient extraction of proteins from recalcitrant plant tissue for subsequent analysis by two‐dimensional gel electrophoresis

This study unveils the single and combined drought and heat impacts on the photosynthetic performance of Coffea arabica cv. Icatu and C. canephora cv. Conilon Clone 153 (CL153). Well-watered (WW) potted plants were gradually submitted to severe water deficit (SWD) along 20 days under adequate temperature (25/20°C, day/night), and thereafter exposed to a gradual temperature rise up to 42/30°C, followed by a 14-day water and temperature recovery. Single drought affected all gas exchanges (including A max) and most fluorescence parameters in both genotypes. However, Icatu maintained F v /F m and RuBisCO activity, and reinforced electron transport rates, carrier contents, and proton gradient regulation (PGR5) and chloroplast NADH dehydrogenase-like (NDH) complex proteins abundance. This suggested negligible non-stomatal limitations of photosynthesis that were accompanied by a triggering of protective cyclic electron transport (CEF) involving both photosystems (PSs). These findings contrasted with declines in RuBisCO and PSs activities, and cytochromes (b 559 , f, b 563) contents in CL153. Remarkable heat tolerance in potential photosynthetic functioning was detected in WW plants of both genotypes (up to 37/28°C or 39/30°C), likely associated with CEF in Icatu. Yet, at 42/30°C the tolerance limit was exceeded. Reduced A max and increased C i

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“…Protein Extraction and Trypsin Proteolysis Protein extraction followed Parkhey et al (2015) with some modifications. Briefly, ca.…”

Section: Proteins Associated With Thylakoid Cyclic Electron Flowmentioning

confidence: 99%

Resilient and Sensitive Key Points of the Photosynthetic Machinery of Coffea spp. to the Single and Superimposed Exposure to Severe Drought and Heat Stresses

et al. 2020

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“…The release of proteins from the microorganisms (Gonzalez‐Balderas, Velasquez‐Orta, Valdez‐Vazquez, & Orta Ledesma, 2020) and the choice of extractant/buffer (Bastida, Jehmlich, Torres, & Garcia, 2018), precipitant, and lysate (Miskiewicz & MacPhee, 2019) have crucial effects on protein extraction. For the extraction of proteins in many substrates, the conditions mentioned above must be optimized systematically (Awad & Brueck, 2019; Parkhey, Chandrakar, Naithani, & Keshavkant, 2015; Wohlbrand et al., 2017), and optimal extraction results had been achieved. For example, for extracting total proteins from the intestine, SDS–containing lysate enables more extensive protein and peptide/proteome identification than urea‐containing lysate (Zhang et al., 2018b).…”

Section: Metaproteomics Analysis Methodsmentioning

confidence: 99%

Metaproteomics insights into traditional fermented foods and beverages

Yang

Fan

2020

Comp Rev Food Sci Food Safe

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Traditional fermented foods and beverages (TFFB) are important dietary components. Multi‐omics techniques have been applied to all aspects of TFFB research to clarify the composition and nutritional value of TFFB, and to reveal the microbial community, microbial interactions, fermentative kinetics, and metabolic profiles during the fermentation process of TFFB. Because of the advantages of metaproteomics in providing functional information, this technology has increasingly been used in research to assess the functional diversity of microbial communities. Metaproteomics is gradually gaining attention in the field of TFFB research because it can reveal the nature of microorganism function at the protein level. This paper reviews the common methods of metaproteomics applied in TFFB research; systematically summarizes the results of metaproteomics research on TFFB, such as sauces, wines, fermented tea, cheese, and fermented fish; and compares the differences in conclusions reached through metaproteomics versus other omics methods. Metaproteomics has great advantages in revealing the microbial functions in TFFB and the interaction between the materials and microbial community. In the future, metaproteomics should be further applied to the study of functional protein markers and protein interaction in TFFB; multi‐omics technology requires further integration to reveal the molecular nature of TFFB fermentation.

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“…photosynthetically active) tissues (5,6). As a result numerous protein extraction methods using various combinations of trichloroacetic acid, acetone, methanol, chloroform, phenol, detergents, and molecular weight cutoff filters have been evaluated on plant tissues (5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22). However, nearly all the evaluations of extraction methods have examined compatibility with 2-dimensional gel electrophoresis rather than reverse phase liquid chromatography (LC), which is necessary for deep and quantitative proteome profiling.…”

Section: Introductionmentioning

confidence: 99%

Assessment and refinement of sample preparation methods for deep and quantitative plant proteome profiling

Song

Hsu

Walley

2018

Preprint

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A major challenge in the field of proteomics is obtaining high quality peptides for comprehensive proteome profiling by liquid chromatography mass spectrometry for many organisms. Here we evaluate and modify a range of sample preparation methods using photosynthetically active Arabidopsis leaf tissues from several developmental timepoints. We find that inclusion of FASP-based on filter digestion improves all protein extraction methods tested. Ultimately, we show that a detergent-free urea-FASP approach enables deep and robust quantification of leaf proteomes. For example, from 4-day-old leaf tissue we profiled up to 11,690 proteins from a single sample replicate. This method should be broadly applicable to researchers working on difficult to process samples from a range of plant and non-plant organisms.

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Efficient extraction of proteins from recalcitrant plant tissue for subsequent analysis by two‐dimensional gel electrophoresis

Cited by 12 publications

References 31 publications

Resilient and Sensitive Key Points of the Photosynthetic Machinery of Coffea spp. to the Single and Superimposed Exposure to Severe Drought and Heat Stresses

Resilient and Sensitive Key Points of the Photosynthetic Machinery of Coffea spp. to the Single and Superimposed Exposure to Severe Drought and Heat Stresses

Metaproteomics insights into traditional fermented foods and beverages

Assessment and refinement of sample preparation methods for deep and quantitative plant proteome profiling

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