The sequence 581Ser-610Val in the fibrinogen Aα chain is responsible for the formation of complexes between plasminogen and αC-regions of fibrin(ogen)
Objective: This study aimed to identify the binding sites for plasminogen (Pg) and its kringle-containing fragments within the αC-region of fibrin(ogen). This investigation is crucial while the conversion of fibrinogen into fibrin induces conformational changes that expose binding sites for Pg and t...
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Elsevier
2024-12-01
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| Series: | Heliyon |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405844024168831 |
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| author | Lada Kapustianenko Tetiana Grinenko Andrew Rebriev Artem Tykhomyrov |
| author_facet | Lada Kapustianenko Tetiana Grinenko Andrew Rebriev Artem Tykhomyrov |
| author_sort | Lada Kapustianenko |
| collection | DOAJ |
| description | Objective: This study aimed to identify the binding sites for plasminogen (Pg) and its kringle-containing fragments within the αC-region of fibrin(ogen). This investigation is crucial while the conversion of fibrinogen into fibrin induces conformational changes that expose binding sites for Pg and tissue-type Pg activator (tPA), facilitating effective zymogen activation on the fibrin surface. Methods: Two C-terminal fragments of the Aα chain ‒ 45 kDa (225Val-610Val) and 40 kDa (225Val-580Lys), were obtained through plasmin hydrolysis of human fibrinogen and subsequently characterized using MALDI TOF mass spectrometry. The interactions of Glu-Pg and Lys-Pg, as well as Pg kringle fragments (K1-3, K4, and K5), with the obtained αC truncated polypeptides were analyzed using ELISA and Western blot techniques with the use of specific antibodies. Results: It was demonstrated that Pg and its fragments K1-3, K4, and K5 interact exclusively with the 45-kDa fragment (225Val-610Val) of the αC region of fibrinogen with high affinity in a concentration-dependent manner (Kd values for Glu-Pg = 7.10 × 10−9 M, Lys-Pg = 6.01 × 10−9 M, K1-3 = 1.08 × 10−7 M, K4 = 5.06 × 10−7 M, and K5 = 2.50 × 10−7 M). This fragment, unlike the 40-kDa polypeptide (225Val-580Lys), contains the α581Ser-610Val sequence. Conclusions: It was shown that the sequence 581Ser-610Val of fibrinogen Aα-chain, which becomes exposed during the conversion of fibrinogen to fibrin, is essential for the formation of complexes between Pg and αC regions of fibrin(ogen), thereby contributing to the initiation and regulation of fibrinolysis. |
| format | Article |
| id | doaj-art-17e80c7baaa24d12bfc5bc6c959858d1 |
| institution | OA Journals |
| issn | 2405-8440 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Elsevier |
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| series | Heliyon |
| spelling | doaj-art-17e80c7baaa24d12bfc5bc6c959858d12025-08-20T02:38:10ZengElsevierHeliyon2405-84402024-12-011023e4085210.1016/j.heliyon.2024.e40852The sequence 581Ser-610Val in the fibrinogen Aα chain is responsible for the formation of complexes between plasminogen and αC-regions of fibrin(ogen)Lada Kapustianenko0Tetiana Grinenko1Andrew Rebriev2Artem Tykhomyrov3Corresponding author.; Palladin Institute of Biochemistry of NAS of Ukraine, Kyiv, UkrainePalladin Institute of Biochemistry of NAS of Ukraine, Kyiv, UkrainePalladin Institute of Biochemistry of NAS of Ukraine, Kyiv, UkraineCorresponding author.; Palladin Institute of Biochemistry of NAS of Ukraine, Kyiv, UkraineObjective: This study aimed to identify the binding sites for plasminogen (Pg) and its kringle-containing fragments within the αC-region of fibrin(ogen). This investigation is crucial while the conversion of fibrinogen into fibrin induces conformational changes that expose binding sites for Pg and tissue-type Pg activator (tPA), facilitating effective zymogen activation on the fibrin surface. Methods: Two C-terminal fragments of the Aα chain ‒ 45 kDa (225Val-610Val) and 40 kDa (225Val-580Lys), were obtained through plasmin hydrolysis of human fibrinogen and subsequently characterized using MALDI TOF mass spectrometry. The interactions of Glu-Pg and Lys-Pg, as well as Pg kringle fragments (K1-3, K4, and K5), with the obtained αC truncated polypeptides were analyzed using ELISA and Western blot techniques with the use of specific antibodies. Results: It was demonstrated that Pg and its fragments K1-3, K4, and K5 interact exclusively with the 45-kDa fragment (225Val-610Val) of the αC region of fibrinogen with high affinity in a concentration-dependent manner (Kd values for Glu-Pg = 7.10 × 10−9 M, Lys-Pg = 6.01 × 10−9 M, K1-3 = 1.08 × 10−7 M, K4 = 5.06 × 10−7 M, and K5 = 2.50 × 10−7 M). This fragment, unlike the 40-kDa polypeptide (225Val-580Lys), contains the α581Ser-610Val sequence. Conclusions: It was shown that the sequence 581Ser-610Val of fibrinogen Aα-chain, which becomes exposed during the conversion of fibrinogen to fibrin, is essential for the formation of complexes between Pg and αC regions of fibrin(ogen), thereby contributing to the initiation and regulation of fibrinolysis.http://www.sciencedirect.com/science/article/pii/S2405844024168831PlasminogenKringle fragmentstPAFibrin(ogen)αC-regionMALDI-TOF mass spectrometry |
| spellingShingle | Lada Kapustianenko Tetiana Grinenko Andrew Rebriev Artem Tykhomyrov The sequence 581Ser-610Val in the fibrinogen Aα chain is responsible for the formation of complexes between plasminogen and αC-regions of fibrin(ogen) Heliyon Plasminogen Kringle fragments tPA Fibrin(ogen) αC-region MALDI-TOF mass spectrometry |
| title | The sequence 581Ser-610Val in the fibrinogen Aα chain is responsible for the formation of complexes between plasminogen and αC-regions of fibrin(ogen) |
| title_full | The sequence 581Ser-610Val in the fibrinogen Aα chain is responsible for the formation of complexes between plasminogen and αC-regions of fibrin(ogen) |
| title_fullStr | The sequence 581Ser-610Val in the fibrinogen Aα chain is responsible for the formation of complexes between plasminogen and αC-regions of fibrin(ogen) |
| title_full_unstemmed | The sequence 581Ser-610Val in the fibrinogen Aα chain is responsible for the formation of complexes between plasminogen and αC-regions of fibrin(ogen) |
| title_short | The sequence 581Ser-610Val in the fibrinogen Aα chain is responsible for the formation of complexes between plasminogen and αC-regions of fibrin(ogen) |
| title_sort | sequence 581ser 610val in the fibrinogen aα chain is responsible for the formation of complexes between plasminogen and αc regions of fibrin ogen |
| topic | Plasminogen Kringle fragments tPA Fibrin(ogen) αC-region MALDI-TOF mass spectrometry |
| url | http://www.sciencedirect.com/science/article/pii/S2405844024168831 |
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