Binding of the kringle-2 domain of human plasminogen to streptococcal PAM-type M-protein causes dissociation of PAM dimers
© 2021 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd..
The direct binding of human plasminogen (hPg), via its kringle-2 domain (K2hPg ), to streptococcal M-protein (PAM), largely contributes to the pathogenesis of Pattern D Group A Streptococcus pyogenes (GAS). However, the mechanism of complex formation is unknown. In a system consisting of a Class II PAM from Pattern D GAS isolate NS88.2 (PAMNS88.2 ), with one K2hPg binding a-repeat in its A-domain, we employed biophysical techniques to analyze the mechanism of the K2hPg /PAMNS88.2 interaction. We show that apo-PAMNS88.2 is a coiled-coil homodimer (M.Wt. ~80 kDa) at 4°C-25°C, and is monomeric (M.Wt. ~40 kDa) at 37°C, demonstrating a temperature-dependent dissociation of PAMNS88.2 over a narrow temperature range. PAMNS88.2 displayed a single tight binding site for K2hPg at 4°C, which progressively increased at 25°C through 37°C. We isolated the K2hPg /PAMNS88.2 complexes at 4°C, 25°C, and 37°C and found molecular weights of ~50 kDa at each temperature, corresponding to a 1:1 (m:m) K2hPg /PAMNS88.2 monomer complex. hPg activation experiments by streptokinase demonstrated that the hPg/PAMNS88.2 monomer complexes are fully functional. The data show that PAM dimers dissociate into functional monomers at physiological temperatures or when presented with the active hPg module (K2hPg ) showing that PAM is a functional monomer at 37°C.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:10 |
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| Enthalten in: |
MicrobiologyOpen - 10(2021), 6 vom: 21. Nov., Seite e1252 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Ayinuola, Olawole [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 28.02.2022 Date Revised 04.04.2024 published: Print Citation Status MEDLINE |
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| doi: |
10.1002/mbo3.1252 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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NLM335001173 |
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| 100 | 1 | |a Ayinuola, Olawole |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Binding of the kringle-2 domain of human plasminogen to streptococcal PAM-type M-protein causes dissociation of PAM dimers |
| 264 | 1 | |c 2021 | |
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| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2021 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd. | ||
| 520 | |a The direct binding of human plasminogen (hPg), via its kringle-2 domain (K2hPg ), to streptococcal M-protein (PAM), largely contributes to the pathogenesis of Pattern D Group A Streptococcus pyogenes (GAS). However, the mechanism of complex formation is unknown. In a system consisting of a Class II PAM from Pattern D GAS isolate NS88.2 (PAMNS88.2 ), with one K2hPg binding a-repeat in its A-domain, we employed biophysical techniques to analyze the mechanism of the K2hPg /PAMNS88.2 interaction. We show that apo-PAMNS88.2 is a coiled-coil homodimer (M.Wt. ~80 kDa) at 4°C-25°C, and is monomeric (M.Wt. ~40 kDa) at 37°C, demonstrating a temperature-dependent dissociation of PAMNS88.2 over a narrow temperature range. PAMNS88.2 displayed a single tight binding site for K2hPg at 4°C, which progressively increased at 25°C through 37°C. We isolated the K2hPg /PAMNS88.2 complexes at 4°C, 25°C, and 37°C and found molecular weights of ~50 kDa at each temperature, corresponding to a 1:1 (m:m) K2hPg /PAMNS88.2 monomer complex. hPg activation experiments by streptokinase demonstrated that the hPg/PAMNS88.2 monomer complexes are fully functional. The data show that PAM dimers dissociate into functional monomers at physiological temperatures or when presented with the active hPg module (K2hPg ) showing that PAM is a functional monomer at 37°C | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a analytical ultracentrifugation | |
| 650 | 4 | |a circular dichroism | |
| 650 | 4 | |a human plasminogen | |
| 650 | 4 | |a isothermal calorimetry | |
| 650 | 4 | |a kringle domains | |
| 650 | 4 | |a streptococcal plasminogen binding M-protein | |
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| 650 | 7 | |a Bacterial Outer Membrane Proteins |2 NLM | |
| 650 | 7 | |a Carrier Proteins |2 NLM | |
| 650 | 7 | |a streptococcal M protein |2 NLM | |
| 650 | 7 | |a Plasminogen |2 NLM | |
| 650 | 7 | |a 9001-91-6 |2 NLM | |
| 650 | 7 | |a Streptokinase |2 NLM | |
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| 700 | 1 | |a Ayinuola, Yetunde A |e verfasserin |4 aut | |
| 700 | 1 | |a Qiu, Cunjia |e verfasserin |4 aut | |
| 700 | 1 | |a Lee, Shaun W |e verfasserin |4 aut | |
| 700 | 1 | |a Ploplis, Victoria A |e verfasserin |4 aut | |
| 700 | 1 | |a Castellino, Francis J |e verfasserin |4 aut | |
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