| Title |
Comprehensive Characterization of Swine Cardiac Troponin T Proteoforms by Top-Down Mass Spectrometry
|
|---|---|
| Published in |
Journal of the American Society for Mass Spectrometry, April 2018
|
| DOI | 10.1007/s13361-018-1925-y |
| Pubmed ID | |
| Authors |
Ziqing Lin, Fang Guo, Zachery R. Gregorich, Ruixiang Sun, Han Zhang, Yang Hu, Dhanansayan Shanmuganayagam, Ying Ge |
| Abstract |
Cardiac troponin T (cTnT) regulates the Ca2+-mediated interaction between myosin thick filaments and actin thin filaments during cardiac contraction and relaxation. cTnT is released into the blood following injury, and increased serum levels of the protein are used clinically as a biomarker for myocardial infarction. Moreover, mutations in cTnT are causative in a number of familial cardiomyopathies. With the increasing use of large animal (swine) model to recapitulate human diseases, it is essential to characterize species-dependent protein sequence variants, alternative RNA splicing, and post-translational modifications (PTMs), but challenges remain due to the incomplete database and lack of validation of the predicted splicing isoforms. Herein, we integrated top-down mass spectrometry (MS) with online liquid chromatography (LC) and immunoaffinity purification to comprehensively characterize miniature swine cTnT proteoforms, including those arising from alternative RNA splicing and PTMs. A total of seven alternative splicing isoforms of cTnT were identified by LC/MS from swine left ventricular tissue, with each isoform containing un-phosphorylated and mono-phosphorylated proteoforms. The phosphorylation site was localized to Ser1 for the mono-phosphorylated proteoforms of cTnT1, 3, 4, and 6 by online MS/MS combining collisionally activated dissociation (CAD) and electron transfer dissociation (ETD). Offline MS/MS on Fourier-transform ion cyclotron resonance (FT-ICR) mass spectrometer with CAD and electron capture dissociation (ECD) was then utilized to achieve deep sequencing of mono-phosphorylated cTnT1 (35.2 kDa) with a high sequence coverage of 87%. Taken together, this study demonstrated the unique advantage of top-down MS in the comprehensive characterization of protein alternative splicing isoforms together with PTMs. Graphical Abstract ᅟ. |
X Demographics
The data shown below were collected from the profiles of 4 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 | 2 | 50% |
| United Kingdom | 1 | 25% |
| Unknown | 1 | 25% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Scientists | 2 | 50% |
| Science communicators (journalists, bloggers, editors) | 1 | 25% |
| Members of the public | 1 | 25% |
Mendeley readers
The data shown below were compiled from readership statistics for 23 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 23 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 5 | 22% |
| Student > Bachelor | 3 | 13% |
| Other | 2 | 9% |
| Researcher | 2 | 9% |
| Professor | 1 | 4% |
| Other | 3 | 13% |
| Unknown | 7 | 30% |
| Readers by discipline | Count | As % |
|---|---|---|
| Chemistry | 9 | 39% |
| Biochemistry, Genetics and Molecular Biology | 2 | 9% |
| Unspecified | 1 | 4% |
| Materials Science | 1 | 4% |
| Medicine and Dentistry | 1 | 4% |
| Other | 0 | 0% |
| Unknown | 9 | 39% |
Attention Score in Context
This research output has an Altmetric Attention Score of 2. 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 19 April 2018.
All research outputs
#15,175,718
of 25,382,440 outputs
Outputs from Journal of the American Society for Mass Spectrometry
#2,291
of 3,835 outputs
Outputs of similar age
#182,008
of 343,375 outputs
Outputs of similar age from Journal of the American Society for Mass Spectrometry
#33
of 77 outputs
Altmetric has tracked 25,382,440 research outputs across all sources so far. This one is in the 38th percentile – i.e., 38% of other outputs scored the same or lower than it.
So far Altmetric has tracked 3,835 research outputs from this source. They receive a mean Attention Score of 3.8. This one is in the 38th percentile – i.e., 38% of its peers scored the same or lower than it.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 343,375 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 45th percentile – i.e., 45% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 77 others from the same source and published within six weeks on either side of this one. This one has gotten more attention than average, scoring higher than 55% of its contemporaries.