Identification of the Binding Surface on β-Lactamase for GroEL by Limited Proteolysis and MALDI-Mass Spectrometry†

Biochemistry ◽  
1998 ◽  
Vol 37 (33) ◽  
pp. 11660-11669 ◽  
Author(s):  
Pietro Gervasoni ◽  
Werner Staudenmann ◽  
Peter James ◽  
Andreas Plückthun
Catalysts ◽  
2019 ◽  
Vol 9 (10) ◽  
pp. 833 ◽  
Author(s):  
Rosulek ◽  
Darebna ◽  
Pompach ◽  
Slavata ◽  
Novak

A large number of different enzyme immobilization techniques are used in the field of life sciences, clinical diagnostics, or biotechnology. Most of them are based on a chemically mediated formation of covalent bond between an enzyme and support material. The covalent bond formation is usually associated with changes of the enzymes’ three-dimensional structure that can lead to reduction of enzyme activity. The present work demonstrates a potential of an ambient ion-landing technique to effectively immobilize enzymes on conductive supports for direct matrix-assisted laser desorption/ionization (MALDI) mass spectrometry analyses of reaction products. Ambient ion landing is an electrospray-based technique allowing strong and stable noncovalent and nondestructive enzyme deposition onto conductive supports. Three serine proteolytic enzymes including trypsin, α-chymotrypsin, and subtilisin A were immobilized onto conductive indium tin oxide glass slides compatible with MALDI mass spectrometry. The functionalized MALDI chips were used for in situ time-limited proteolysis of proteins and protein–ligand complexes to monitor their structural changes under different conditions. The data from limited proteolysis using MALDI chips fits to known or predicted protein structures. The results show that functionalized MALDI chips are sensitive, robust, and fast and might be automated for general use in the field of structural biology.


2018 ◽  
Author(s):  
Shazia Khan ◽  
Diego F Cobice ◽  
Dawn EW Livingstone ◽  
C Logan Mackay ◽  
Scott P Webster ◽  
...  

2020 ◽  
Vol 75 (14) ◽  
pp. 1749-1757
Author(s):  
E. A. Anikeenko ◽  
E. N. Rakhmatullina ◽  
D. I. Falev ◽  
O. Yu. Khoroshev ◽  
N. V. Ul’yanovskii ◽  
...  

Author(s):  
Riccardo Zecchi ◽  
Pietro Franceschi ◽  
Laura Tigli ◽  
Davide Amidani ◽  
Chiara Catozzi ◽  
...  

AbstractCorticosteroids as budesonide can be effective in reducing topic inflammation processes in different organs. Therapeutic use of budesonide in respiratory diseases, like asthma, chronic obstructive pulmonary disease, and allergic rhinitis is well known. However, the pulmonary distribution of budesonide is not well understood, mainly due to the difficulties in tracing the molecule in lung samples without the addition of a label. In this paper, we present a matrix-assisted laser desorption/ionization mass spectrometry imaging protocol that can be used to visualize the pulmonary distribution of budesonide administered to a surfactant-depleted adult rabbit. Considering that budesonide is not easily ionized by MALDI, we developed an on-tissue derivatization method with Girard’s reagent P followed by ferulic acid deposition as MALDI matrix. Interestingly, this sample preparation protocol results as a very effective strategy to raise the sensitivity towards not only budesonide but also other corticosteroids, allowing us to track its distribution and quantify the drug inside lung samples. Graphical abstract


Biochimie ◽  
2021 ◽  
Vol 185 ◽  
pp. 87-95
Author(s):  
Aleksandr S. Taraskin ◽  
Konstantin K. Semenov ◽  
Aleksandr V. Protasov ◽  
Alexey A. Lozhkov ◽  
Aleksandr A. Tyulin ◽  
...  

2020 ◽  
Vol 58 (6) ◽  
pp. 883-896 ◽  
Author(s):  
Muhammad Zubair Israr ◽  
Dennis Bernieh ◽  
Andrea Salzano ◽  
Shabana Cassambai ◽  
Yoshiyuki Yazaki ◽  
...  

AbstractBackgroundMatrix-assisted laser desorption ionisation (MALDI) mass spectrometry (MS) has been used for more than 30 years. Compared with other analytical techniques, it offers ease of use, high throughput, robustness, cost-effectiveness, rapid analysis and sensitivity. As advantages, current clinical techniques (e.g. immunoassays) are unable to directly measure the biomarker; rather, they measure secondary signals. MALDI-MS has been extensively researched for clinical applications, and it is set for a breakthrough as a routine tool for clinical diagnostics.ContentThis review reports on the principles of MALDI-MS and discusses current clinical applications and the future clinical prospects for MALDI-MS. Furthermore, the review assesses the limitations currently experienced in clinical assays, the advantages and the impact of MALDI-MS to transform clinical laboratories.SummaryMALDI-MS is widely used in clinical microbiology for the screening of microbial isolates; however, there is scope to apply MALDI-MS in the diagnosis, prognosis, therapeutic drug monitoring and biopsy imaging in many diseases.OutlookThere is considerable potential for MALDI-MS in clinic as a tool for screening, profiling and imaging because of its high sensitivity and specificity over alternative techniques.


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