| Chapter title |
Targeted Approach for Proteomic Analysis of a Hidden Membrane Protein
|
|---|---|
| Chapter number | 12 |
| Book title |
Serum/Plasma Proteomics
|
| Published in |
Methods in molecular biology, July 2017
|
| DOI | 10.1007/978-1-4939-7057-5_12 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7056-8, 978-1-4939-7057-5
|
| Authors |
Tania Martins-Marques, Sandra I. Anjo, Teresa Ribeiro-Rodrigues, Bruno Manadas, Henrique Girao |
| Editors |
David W. Greening, Richard J. Simpson |
| Abstract |
Given the properties of plasma membrane proteins, namely, their hydrophobicity, low solubility, and high resistance to digestion and extraction, their identification by traditional mass spectrometry (MS) has been a challenging task. Hence, proteomic studies involving the transmembrane protein connexin43 (Cx43) are scarce. Additionally, studies demonstrating the presence of proteins embedded in the lipid bilayer of extracellular vesicles (EVs) are difficult to perform and require specific changes and fine adjustments in the experimental and technical procedure to allow their detection by MS. In this review, we provide a detailed description of the protocol we have used to detect Cx43 in EVs of human peripheral blood. This includes some of the modifications that we have introduced in order to improve the detection of Cx43 in EVs, including an optimization of vesicle isolation, Cx43 purification, MS acquisition data, and further analysis. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 1 | 50% |
| Unknown | 1 | 50% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 2 | 100% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 50 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 14 | 28% |
| Student > Ph. D. Student | 9 | 18% |
| Student > Bachelor | 7 | 14% |
| Student > Doctoral Student | 4 | 8% |
| Student > Master | 3 | 6% |
| Other | 4 | 8% |
| Unknown | 9 | 18% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 14 | 28% |
| Chemistry | 6 | 12% |
| Medicine and Dentistry | 5 | 10% |
| Engineering | 4 | 8% |
| Agricultural and Biological Sciences | 3 | 6% |
| Other | 5 | 10% |
| Unknown | 13 | 26% |