Bacterial Secretion Systems
Springer US
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Attention Score in Context
| Chapter title |
Starting with an Integral Membrane Protein Project for Structural Biology: Production, Purification, Detergent Quantification, and Buffer Optimization-Case Study of the Exporter CntI from Pseudomonas aeruginosa.
|
|---|---|
| Book title |
Bacterial Secretion Systems
|
| Published in |
Methods in molecular biology, January 2024
|
| DOI | 10.1007/978-1-0716-3445-5_26 |
| Pubmed ID | |
| Book ISBNs |
978-1-07-163444-8, 978-1-07-163445-5
|
| Authors |
Mégret-Cavalier, Maxime, Pozza, Alexandre, Cece, Quentin, Bonneté, Françoise, Broutin, Isabelle, Phan, Gilles |
| Abstract |
Production, extraction, purification, and stabilization of integral membrane proteins are key steps for successful structural biology studies, in particular for X-ray crystallography or single particle microscopy. Here, we present the purification protocol of CntI from Pseudomonas aeruginosa, a new metallophore exporter of the Drug Metabolite Transporter (DMT) family involved in pseudopaline secretion. Subsequent to CntI purification, we optimized the buffer pH, salts, and additives by differential scanning fluorimetry (DSF), also known as Thermofluor Assay (TFA) or fluorescent thermal stability assay (FTSA), with the use of dye 1-AnilinoNaphthalene-8-Sulfonic acid (ANS), a fluorescent molecule compatible with detergents. After the buffer optimization, the purified CntI was analyzed by Size Exclusion Chromatography coupled with Multi-Angle Laser Light Scattering (SEC-MALLS), UV absorbance, and Refractive Index detectors, in order to determine the absolute molar mass of the protein-detergent complex, the detergent amount bound to the protein and the amount of protein-free detergent micelles. Altogether, these biophysical techniques give preliminary and mandatory information about the suitability of the purified membrane protein for further biophysical or structural investigations. |
X Demographics
The data shown below were collected from the profiles of 2 X users who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Spain | 1 | 50% |
| United States | 1 | 50% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 2 | 100% |
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 07 November 2023.
All research outputs
#15,633,846
of 24,764,450 outputs
Outputs from Methods in molecular biology
#4,574
of 13,893 outputs
Outputs of similar age
#6,924
of 15,997 outputs
Outputs of similar age from Methods in molecular biology
#6
of 57 outputs
Altmetric has tracked 24,764,450 research outputs across all sources so far. This one is in the 34th percentile – i.e., 34% of other outputs scored the same or lower than it.
So far Altmetric has tracked 13,893 research outputs from this source. They receive a mean Attention Score of 3.5. This one has gotten more attention than average, scoring higher than 63% of its peers.
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 15,997 tracked outputs that were published within six weeks on either side of this one in any source. This one has gotten more attention than average, scoring higher than 53% of its contemporaries.
We're also able to compare this research output to 57 others from the same source and published within six weeks on either side of this one. This one has done well, scoring higher than 84% of its contemporaries.