Evaluation of four protein extraction methods for proteomic analysis of mango peel

Abstract

ABSTRACT. The peel of mango (Mangifera indica L.) is a special plant tissue that contains many compounds that interfere with protein extraction. A successful separation with Two-dimensional electrophoresis (2-DE) is the key step for proteomic analysis. To evaluate the efficiencies of mango peel protein extraction for 2-DE, four extraction methods were tested: 1) 2-D clean-up kit, 2) trichloroacetic acid/acetone precipitation, 3) phenol extraction, 4) phenol with methanol/ammonium acetate precipitation. The results showed that the phenol with methanol/ammonium acetate precipitation produced the best quality protein extraction and separation. Proteins were separated in 30-70 and >70 kDa ranges better than with the other methods. Acidic proteins had better resolution with fewer horizontal and vertical streaks. Sixteen proteins were identified by maxtrix-assisted laser desorption/ ionisation time-of-flight tandem mass spectrometry (MALDI-TOF/ TOF-MS/MS). The result demonstrated that each of these four methods can be used to prepare mango peel proteins. The phenol with methanol/ ammonium acetate precipitation was the best choice for proteomic analysis of mango peel.

Comparing and integrating TMT‐SPS‐MS3 and label‐free quantitative approaches for proteomics scrutiny in recalcitrant Mango (Mangifera indica L.) peel tissue during postharvest period

“…Phe extraction outperforms extraction buffers containing CaCl 2 and LiCl during the extraction of proteins from purified cell wall of skin and seed of ripened grape berry ( Vitis vinifera L. cv. Cabernet Sauvignon grapevines) [31] and had higher quality protein extraction and separation compared to 2‐D clean‐up kit and TCA/acetone precipitation in mango peel [12]. Previous tests have been based on protein yield and analysis with 2D gels coupled to mass spectrometry.…”

“…For example, a previous proteomic study in the peels of different mango cultivars during postharvest period using phenolic extraction, strong cation exchange (SCX) peptide fractionation, and iTRAQ‐SPS‐MS3 in combination with metabolomic analyses showed that they contain nutraceutical molecules [11]. However, mango peel is particularly difficult to analyse, as evidenced by the low number of proteins reported in this tissue when compared to others [12], highlighting the limitations of the current proteomic pipelines used for the analysis of peel tissues so far. Due to the essential role of cuticular layer as modulator of fruit soften during postharvest period, it is paramount to focus efforts on characterizing the proteome of mango peel.…”

Advances in Physiological, Transcriptomic, Proteomic, Metabolomic, and Molecular Genetic Approaches for Enhancing Mango Fruit Quality

“…1') were subjected to proteomic analysis and identified proteins involved in immunity and defense, oxidation and reduction, and amino acid and carbohydrate metabolism. 140 Ultraviolet-C treatment can potentially be used to maintain the fruit quality as well as extend shelf life of mangoes. 136 The effects of UV−C on the quality of fruits at the proteomic level were investigated from mango pulp tissue (cv.…”

“…Fresh ripe mangoes (‘Tainong No. 1’) were subjected to proteomic analysis and identified proteins involved in immunity and defense, oxidation and reduction, and amino acid and carbohydrate metabolism …”

Proteomic Contributions to Medicinal Plant Research: From Plant Metabolism to Pharmacological Action

“…A protein catalogue was provided for the mango plant, which is a leading fruit crop and produces the unique xanthonoid called mangiferin [ 69 , 70 ]. A recent study improved protein purification methods for extracting proteins existing in small quantity in mango fruit pulp and banana [ 71 , 72 ]. In addition to the popular plants, some of the indigenous and underutilized plants were analyzed for their phytochemicals and physiology [ 73 ].…”