Trypsinogen and chymotrypsinogen: the mysterious hyper‐reactivity of selected cysteines is still present after their divergent evolution
| Autor: | Federica Iavarone, Mozhgan Boroumand, Giorgio Ricci, Massimo Castagnola, Giada Cattani, Alessio Bocedi, Giorgia Gambardella |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Protein Folding Evolution chymotrypsinogen Trypsinogen Chymotrypsinogen Biochemistry Evolution Molecular molten globule 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Humans Cysteine Disulfides Ribonuclease Settore BIO/10 Settore BIO/10 - BIOCHIMICA Molecular Biology cysteine reactivity oxidative folding trypsinogen Glutathione Oxidation-Reduction biology Oxidative folding Molecular Cell Biology Molten globule Divergent evolution 030104 developmental biology chemistry 030220 oncology & carcinogenesis Biophysics biology.protein Lysozyme |
| Zdroj: | The FEBS Journal. 288:6003-6018 |
| ISSN: | 1742-4658 1742-464X |
| DOI: | 10.1111/febs.15886 |
| Popis: | An enigmatic and never described hyper-reactivity of most of the cysteines resident in the reduced, molten globule-like intermediate of a few proteins has been recently discovered. In particular, all ten cysteines of chymotrypsinogen showed hundred times increased reactivity against hydrophobic reagents. A single cysteine (Cys1) was also found thousand times more reactive toward GSSG, making speculate that a single glutathionylation could represent the primordial event of its oxidative folding. In the present study, we compare these kinetic properties with those present in trypsinogen taken in its reduced, molten globule-like intermediate and identify the origin of these unusual properties. Despite the divergent evolution of these two proteins, the different amount of disulfides and the very different three-dimensional localization of three disulfides, their hyper-reactivity toward hydrophobic thiol reagents and disulfides is very similar. Mass spectrometry identifies two cysteines in trypsinogen, Cys148 and Cys197, 800 times more reactive toward GSSG than an unperturbed protein cysteine. These results point towards a stringent and accurate preservation of these peculiar kinetic properties during a divergent evolution suggesting some important role which at the present can only be hypothesized. Similar extraordinary hyper-reactivity has been found also in albumin, ribonuclease and lysozyme confirming that it cannot be considered a kinetic singularity of a single protein. Interestingly, the very flexible and fluctuating structures like those typical of the molten globule status proves capable of enabling sophisticated actions typical of enzymes like binding to GSSG with relevant specificity and high affinity (KD = 0.4 mM) and accelerating the reaction of its cysteines by thousands of times. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
| Nepřihlášeným uživatelům se plný text nezobrazuje | K zobrazení výsledku je třeba se přihlásit. |