Development and comparative analysis of yeast protein extraction protocols for mass spectrometry

Author(s)

de Almeida, Lauana Fogaca ^[1] ; de Moraes, Leonardo Nazario ^[1] ; dos Santos, Lucilene Delazari ^{[2, 3]} ; Valente, Guilherme Targino ^[1]

Affiliation

^[1] Sao Paulo State Univ UNESP, Dept Bioproc & Biotechnol, Sch Agr, Ave Univ 3780, BR-18610034 Botucatu, SP - Brazil

^[2] Sao Paulo State Univ UNESP, Ctr Study Venoms & Venomous Anim CEVAP, Ave Univ 3780, BR-18610034 Botucatu, SP - Brazil

^[3] Sao Paulo State Univ UNESP, Grad Program Trop Dis, Botucatu Med Sch FMB, Ave Prof Mario Rubens Guimaraes Montenegro, BR-18618687 Botucatu, SP - Brazil

• Document type: Journal article
• Source: Analytical Biochemistry; v. 567, p. 90-95, FEB 15 2019.
• Web of Science Citations: 1

Abstract

Mass spectrometry is the most used method for protein identification and quantification. Here we developed four protein extraction protocols precisely for mass spectrometry, and we compared with other ones already published. The best protocol developed by us consists on a simple extraction solution, a heat-shock step, and does not use protease inhibitor; moreover, it is the most efficient and uniform among replicates, besides to be safe, cheap and fast. That method also provided the highest number of proteins uniquely identified and allows finding a diversity of protein classes, which their absence is a problem to be avoided. (AU)

• FAPESP's process: 15/12093-9 - Integrative analysis applied to ethanol tolerance in Saccharomyces cerevisiae strains: an approach using transcriptomes, proteomes, system biology and machine learning
• Grantee: Guilherme Targino Valente
• Support Opportunities: Program for Research on Bioenergy (BIOEN) - Regular Program Grants