| Title |
SPRi-MALDI MS: characterization and identification of a kinase from cell lysate by specific interaction with different designed ankyrin repeat proteins
|
|---|---|
| Published in |
Analytical & Bioanalytical Chemistry, December 2016
|
| DOI | 10.1007/s00216-016-0127-3 |
| Pubmed ID | |
| Authors |
Ulrike Anders, Jonas V. Schaefer, Fatima-Ezzahra Hibti, Chiraz Frydman, Detlev Suckau, Andreas Plückthun, Renato Zenobi |
| Abstract |
We report on the direct coupling of surface plasmon resonance imaging (SPRi) with matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) for the investigation of specific, non-covalent interactions, using the example of designed ankyrin repeat proteins (DARPins) and ribosomal protein S6 kinase 2 (RPS6KA2) directly from lysate of SH-SY5Y cells, derived from human bone marrow. Due to an array format, tracing of binding kinetics of numerous DARPins simultaneously and in real time becomes possible. By optimizing both the proteolytic digest directly on the SPRi chip (amount of trypsin, incubation time, and temperature) as well as the MALDI matrix application (concentration of matrix and number of spray cycles), we are able to identify the specific interaction with RPS6KA2 directly from the cell lysate at a surface coverage of only 0.8 fmol/mm(2). Graphical Abstract Workflow of the direct coupling of SPRi with MALDI mass spectrometry. |
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 | 2 | 67% |
| Unknown | 1 | 33% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Scientists | 2 | 67% |
| Members of the public | 1 | 33% |
Mendeley readers
The data shown below were compiled from readership statistics for 21 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 1 | 5% |
| Unknown | 20 | 95% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 5 | 24% |
| Student > Ph. D. Student | 4 | 19% |
| Other | 1 | 5% |
| Professor | 1 | 5% |
| Student > Doctoral Student | 1 | 5% |
| Other | 2 | 10% |
| Unknown | 7 | 33% |
| Readers by discipline | Count | As % |
|---|---|---|
| Chemistry | 6 | 29% |
| Biochemistry, Genetics and Molecular Biology | 4 | 19% |
| Agricultural and Biological Sciences | 2 | 10% |
| Medicine and Dentistry | 1 | 5% |
| Neuroscience | 1 | 5% |
| Other | 1 | 5% |
| Unknown | 6 | 29% |
Attention Score in Context
This research output has an Altmetric Attention Score of 3. 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 31 July 2017.
All research outputs
#14,403,185
of 25,394,764 outputs
Outputs from Analytical & Bioanalytical Chemistry
#4,289
of 9,624 outputs
Outputs of similar age
#211,436
of 421,540 outputs
Outputs of similar age from Analytical & Bioanalytical Chemistry
#32
of 180 outputs
Altmetric has tracked 25,394,764 research outputs across all sources so far. This one is in the 42nd percentile – i.e., 42% of other outputs scored the same or lower than it.
So far Altmetric has tracked 9,624 research outputs from this source. They receive a mean Attention Score of 3.1. This one has gotten more attention than average, scoring higher than 55% of its peers.
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We're also able to compare this research output to 180 others from the same source and published within six weeks on either side of this one. This one has done well, scoring higher than 81% of its contemporaries.