Label-Free Quantitative Analysis of Mitochondrial Proteomes Using the 
Multienzyme Digestion-Filter Aided Sample Preparation (MED-FASP) and “Total 
Protein Approach”

Wisniewski, Jacek R.

doi:10.1007/978-1-4939-6824-4_6

Datensatz

DATENSATZ AKTIONENEXPORT

Zur Ablage hinzufügen

Lokale TagsFreigabegeschichteDetailsÜbersicht

Freigegeben

Buchkapitel

Label-Free Quantitative Analysis of Mitochondrial Proteomes Using the Multienzyme Digestion-Filter Aided Sample Preparation (MED-FASP) and “Total Protein Approach”

MPG-Autoren

/persons/resource/persons78895

Wisniewski, Jacek R.
Mann, Matthias / Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Max Planck Society;

Externe Ressourcen

Es sind keine externen Ressourcen hinterlegt

Volltexte (beschränkter Zugriff)

Für Ihren IP-Bereich sind aktuell keine Volltexte freigegeben.

Volltexte (frei zugänglich)

Es sind keine frei zugänglichen Volltexte in PuRe verfügbar

Ergänzendes Material (frei zugänglich)

Es sind keine frei zugänglichen Ergänzenden Materialien verfügbar

Zitation

Wisniewski, J. R. (2017). Label-Free Quantitative Analysis of Mitochondrial Proteomes Using the Multienzyme Digestion-Filter Aided Sample Preparation (MED-FASP) and “Total Protein Approach”. In D. Mokranjac, & F. Perocchi (Eds.), Mitochondria (pp. 69-77). New York, NY: Humana Press.

Zitierlink: https://hdl.handle.net/11858/00-001M-0000-002E-80F2-B

Zusammenfassung

Determination of proteome composition and measuring of changes in protein titers provide important information with a substantial value for studying mitochondria. This chapter describes a workflow for the quantitative analysis of mitochondrial proteome with a focus on sample preparation and quantitative analysis of the data. The workflow involves the multienzyme digestion-filter aided sample preparation (MED-FASP) protocol enabling efficient extraction of proteins and high rate of protein-to-peptide conversion. Consecutive protein digestion with Lys C and trypsin enables generation of peptide fractions with minimal overlap, largely increases the number of identified proteins, and extends their sequence coverage. Abundances of proteins identified by multiple peptides can be assessed by the "Total Protein Approach."