title: Evaluation of four protein extraction methods for proteomic analysis of mango peel.
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.
authors.affiliation: State Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China.
authors.forename: D J
authors.initials: DJ
authors.lastname: Liao
authors.affiliation: State Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China.
authors.forename: X P
authors.initials: XP
authors.lastname: Lu
authors.affiliation: State Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China.
authors.forename: H S
authors.initials: HS
authors.lastname: Chen
authors.affiliation: Chinese Academy of Tropical Agricultural Sciences Environment and Plant Protection Institute, Danzhou, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Lu
authors.affiliation: State Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China moziyao@gird.cn.
authors.forename: Z Y
authors.initials: ZY
authors.lastname: Mo
mesh_terms: D000085:Acetates / Q000737:chemistry; D011232:Chemical Precipitation; D005638:Fruit / Q000737:chemistry*; D031022:Mangifera / Q000737:chemistry*; D000432:Methanol / Q000737:chemistry; D019800:Phenol / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification*; D020543:Proteome / Q000737:chemistry / Q000302:isolation & purification*; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry
chemical_list: D000085:Acetates; D010940:Plant Proteins; D020543:Proteome; D019800:Phenol; C018824:ammonium acetate; D000432:Methanol