| Chapter title |
Label-Free Quantitative Analysis of Mitochondrial Proteomes Using the Multienzyme Digestion-Filter Aided Sample Preparation (MED-FASP) and “Total Protein Approach”
|
|---|---|
| Chapter number | 6 |
| Book title |
Mitochondria
|
| Published in |
Methods in molecular biology, March 2017
|
| DOI | 10.1007/978-1-4939-6824-4_6 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6822-0, 978-1-4939-6824-4
|
| Authors |
Jacek R. Wiśniewski |
| Editors |
Dejana Mokranjac, Fabiana Perocchi |
| Abstract |
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." |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 13 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Master | 3 | 23% |
| Professor > Associate Professor | 2 | 15% |
| Student > Ph. D. Student | 2 | 15% |
| Unspecified | 1 | 8% |
| Researcher | 1 | 8% |
| Other | 1 | 8% |
| Unknown | 3 | 23% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 4 | 31% |
| Unspecified | 1 | 8% |
| Agricultural and Biological Sciences | 1 | 8% |
| Computer Science | 1 | 8% |
| Medicine and Dentistry | 1 | 8% |
| Other | 1 | 8% |
| Unknown | 4 | 31% |