| Title |
Comprehensive Proteoform Characterization of Plasma Complement Component C8αβγ by Hybrid Mass Spectrometry Approaches
|
|---|---|
| Published in |
Journal of the American Society for Mass Spectrometry, March 2018
|
| DOI | 10.1007/s13361-018-1901-6 |
| Pubmed ID | |
| Authors |
Vojtech Franc, Jing Zhu, Albert J. R. Heck |
| Abstract |
The human complement hetero-trimeric C8αβγ (C8) protein assembly (~ 150 kDa) is an important component of the membrane attack complex (MAC). C8 initiates membrane penetration and coordinates MAC pore formation. Here, we charted in detail the structural micro-heterogeneity within C8, purified from human plasma, combining high-resolution native mass spectrometry and (glyco)peptide-centric proteomics. The intact C8 proteoform profile revealed at least ~ 20 co-occurring MS signals. Additionally, we employed ion exchange chromatography to separate purified C8 into four distinct fractions. Their native MS analysis revealed even more detailed structural micro-heterogeneity on C8. Subsequent peptide-centric analysis, by proteolytic digestion of C8 and LC-MS/MS, provided site-specific quantitative profiles of different types of C8 glycosylation. Combining all this data provides a detailed specification of co-occurring C8 proteoforms, including experimental evidence on N-glycosylation, C-mannosylation, and O-glycosylation. In addition to the known N-glycosylation sites, two more N-glycosylation sites were detected on C8. Additionally, we elucidated the stoichiometry of all C-mannosylation sites in all the thrombospondin-like (TSP) domains of C8α and C8β. Lastly, our data contain the first experimental evidence of O-linked glycans located on C8γ. Albeit low abundant, these O-glycans are the first PTMs ever detected on this subunit. By placing the observed PTMs in structural models of free C8 and C8 embedded in the MAC, it may be speculated that some of the newly identified modifications may play a role in the MAC formation. Graphical Abstract ᅟ. |
X Demographics
The data shown below were collected from the profiles of 3 X users who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 1 | 33% |
| Unknown | 2 | 67% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Scientists | 1 | 33% |
| Members of the public | 1 | 33% |
| Science communicators (journalists, bloggers, editors) | 1 | 33% |
Mendeley readers
The data shown below were compiled from readership statistics for 40 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 40 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 11 | 28% |
| Researcher | 10 | 25% |
| Student > Bachelor | 3 | 8% |
| Professor > Associate Professor | 3 | 8% |
| Student > Master | 3 | 8% |
| Other | 6 | 15% |
| Unknown | 4 | 10% |
| Readers by discipline | Count | As % |
|---|---|---|
| Chemistry | 9 | 23% |
| Biochemistry, Genetics and Molecular Biology | 7 | 18% |
| Agricultural and Biological Sciences | 7 | 18% |
| Pharmacology, Toxicology and Pharmaceutical Science | 4 | 10% |
| Medicine and Dentistry | 2 | 5% |
| Other | 6 | 15% |
| Unknown | 5 | 13% |
Attention Score in Context
This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 27 March 2018.
All research outputs
#17,292,294
of 25,382,440 outputs
Outputs from Journal of the American Society for Mass Spectrometry
#2,724
of 3,835 outputs
Outputs of similar age
#226,541
of 350,479 outputs
Outputs of similar age from Journal of the American Society for Mass Spectrometry
#42
of 73 outputs
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