Quench me if you can: Alpha-2-macroglobulin trypsin complexes enable serum biomarker analysis by MALDI mass spectrometry
| Autor: | Aram A. Shaldzhyan, Sergey A. Klotchenko, Konstantin K. Semenov, Olga A. Mirgorodskaya, Edward Ramsay, Alexey A. Lozhkov, Yana A. Zabrodskaya, Aleksandr S. Taraskin, A.V. Protasov, Aleksandr A. Tyulin |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
medicine.medical_treatment Peptide Mass spectrometry Biochemistry alpha-2-Macroglobulin 03 medical and health sciences Hydrolysis Serum biomarkers medicine Humans Trypsin alpha-Macroglobulins chemistry.chemical_classification Protease Chromatography 030102 biochemistry & molecular biology biology General Medicine Blood proteins In vitro 030104 developmental biology chemistry Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization biology.protein Peptides Biomarkers medicine.drug |
| Zdroj: | Biochimie. 185 |
| ISSN: | 1638-6183 |
| Popis: | One of the main functions of alpha-2-macroglobulin (A2M) in human blood serum is the binding of all classes of protease. It is known that trypsin, after such interaction, possesses modified proteolytic activity. Trypsin first hydrolyzes two bonds in A2M’s ‘bait region’, and the peptide705VGFYESDVMGR715is released from A2M. In this work, specifics of the A2M-trypsin interaction were used to determine A2M concentration directly in human blood serum using MALDI mass-spectrometry. Following exogenous addition of trypsin to human blood serumin vitro, the concentration of the VGFYESDVMGR peptide was measured, using its isotopically-labeled analogue (18O), and A2M concentration was calculated. The optimized mass spectrometric approach was verified using a standard method for A2M concentration determination (ELISA) and the relevant statistical analysis methods. It was also shown that trypsin’s modified proteolytic activity in the presence of serum A2M can be used to analyze other serum proteins, including potential biomarkers of pathological processes. Thus, this work describes a promising approach to serum biomarker analysis that can be technically extended in several useful directions. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect