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

Autor(es)

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

Afiliação do(s) autor(es)

^[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

• Tipo de documento: Artigo Científico
• Fonte: Analytical Biochemistry; v. 567, p. 90-95, FEB 15 2019.
• Citações Web of Science: 1

Resumo

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)

• Processo FAPESP: 15/12093-9 - Análises integrativas aplicadas à tolerância ao etanol em linhagens de Saccharomyces cerevisiae: uma abordagem envolvendo transcriptoma, proteômica, biologia de sistemas e aprendizado de máquina
• Beneficiário: Guilherme Targino Valente
• Modalidade de apoio: Auxílio à Pesquisa - Programa BIOEN - Regular