Computer-aided engineering of staphylokinase toward enhanced
affinity and selectivity for plasmin

J 2022

Computer-aided engineering of staphylokinase toward enhanced affinity and selectivity for plasmin

NIKITIN, Dmitri; Jan MIČAN; Martin TOUL; David BEDNÁŘ; Michaela PEŠKOVÁ et al.

Základní údaje

Originální název

Computer-aided engineering of staphylokinase toward enhanced affinity and selectivity for plasmin

Autoři

Vydání

Computational and Structural Biotechnology Journal, Amsterdam, Elsevier, 2022, 2001-0370

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Nizozemské království

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 6.000

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14310/22:00126020

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

Acute myocardial infarction; Stroke treatments; Thrombolytics; Plasminogen activators; Staphylokinase; Rational design; Affinity engineering; Enzyme kinetics; AffiLib

Štítky

14110127, 14110512, 14313050, 14314010, 143180, CF BIC, CF NANO, podil, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 21. 2. 2025 15:40, Mgr. Michaela Hylsová, Ph.D.

Anotace

V originále

Cardio-and cerebrovascular diseases are leading causes of death and disability, resulting in one of the highest socio-economic burdens of any disease type. The discovery of bacterial and human plasminogen activators and their use as thrombolytic drugs have revolutionized treatment of these pathologies. Fibrin specific agents have an advantage over non-specific factors because of lower rates of deleterious side effects. Specifically, staphylokinase (SAK) is a pharmacologically attractive indirect plasminogen activator protein of bacterial origin that forms stoichiometric noncovalent complexes with plasmin, promoting the conversion of plasminogen into plasmin. Here we report a computer-assisted re-design of the molecular surface of SAK to increase its affinity for plasmin. A set of computationally designed SAK mutants was produced recombinantly and biochemically characterized. Screening revealed a pharmacologically interesting SAK mutant with-7-fold enhanced affinity toward plasmin,-10-fold improved plasmin selectivity and moderately higher plasmin-generating efficiency in vitro. Collectively, the results obtained provide a framework for SAK engineering using computational affinity-design that could pave the way to next-generation of effective, highly selective, and less toxic thrombolytics.

Návaznosti

LM2018131, projekt VaV

Název: Česká národní infrastruktura pro biologická data (Akronym: ELIXIR-CZ)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Česká národní infrastruktura pro biologická data

LM2018140, projekt VaV

Název: e-Infrastruktura CZ (Akronym: e-INFRA CZ)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, e-Infrastruktura CZ

90127, velká výzkumná infrastruktura

Název: CIISB II

Přiložené soubory

1860499.pdf

Citovat

NIKITIN, Dmitri; Jan MIČAN; Martin TOUL; David BEDNÁŘ; Michaela PEŠKOVÁ; Patrícia KITTOVÁ; Sandra THALEROVÁ; Jan VÍTEČEK; Jiří DAMBORSKÝ; Robert MIKULÍK; Sarel J. FLEISHMAN; Zbyněk PROKOP a Martin MAREK. Computer-aided engineering of staphylokinase toward enhanced affinity and selectivity for plasmin. Computational and Structural Biotechnology Journal. Amsterdam: Elsevier, 2022, roč. 20, May 2022, s. 1366-1377. ISSN 2001-0370. Dostupné z: https://doi.org/10.1016/j.csbj.2022.03.004.

@article{1860499,
   author = {Nikitin, Dmitri and Mičan, Jan and Toul, Martin and Bednář, David and Pešková, Michaela and Kittová, Patrícia and Thalerová, Sandra and Víteček, Jan and Damborský, Jiří and Mikulík, Robert and Fleishman, Sarel J. and Prokop, Zbyněk and Marek, Martin},
   article_location = {Amsterdam},
   article_number = {May 2022},
   doi = {https://doi.org/10.1016/j.csbj.2022.03.004},
   keywords = {Acute myocardial infarction; Stroke treatments; Thrombolytics; Plasminogen activators; Staphylokinase; Rational design; Affinity engineering; Enzyme kinetics; AffiLib},
   language = {eng},
   issn = {2001-0370},
   journal = {Computational and Structural Biotechnology Journal},
   title = {Computer-aided engineering of staphylokinase toward enhanced affinity and selectivity for plasmin},
   url = {https://www.sciencedirect.com/science/article/pii/S2001037022000794?via%3Dihub},
   volume = {20},
   year = {2022}
}

TY  - JOUR
ID  - 1860499
AU  - Nikitin, Dmitri - Mičan, Jan - Toul, Martin - Bednář, David - Pešková, Michaela - Kittová, Patrícia - Thalerová, Sandra - Víteček, Jan - Damborský, Jiří - Mikulík, Robert - Fleishman, Sarel J. - Prokop, Zbyněk - Marek, Martin
PY  - 2022
TI  - Computer-aided engineering of staphylokinase toward enhanced affinity and selectivity for plasmin
JF  - Computational and Structural Biotechnology Journal
VL  - 20
IS  - May 2022
SP  - 1366-1377
EP  - 1366-1377
PB  - Elsevier
SN  - 20010370
KW  - Acute myocardial infarction
KW  - Stroke treatments
KW  - Thrombolytics
KW  - Plasminogen activators
KW  - Staphylokinase
KW  - Rational design
KW  - Affinity engineering
KW  - Enzyme kinetics
KW  - AffiLib
UR  - https://www.sciencedirect.com/science/article/pii/S2001037022000794?via%3Dihub
N2  - Cardio-and cerebrovascular diseases are leading causes of death and disability, resulting in one of the highest socio-economic burdens of any disease type. The discovery of bacterial and human plasminogen activators and their use as thrombolytic drugs have revolutionized treatment of these pathologies. Fibrin specific agents have an advantage over non-specific factors because of lower rates of deleterious side effects. Specifically, staphylokinase (SAK) is a pharmacologically attractive indirect plasminogen activator protein of bacterial origin that forms stoichiometric noncovalent complexes with plasmin, promoting the conversion of plasminogen into plasmin. Here we report a computer-assisted re-design of the molecular surface of SAK to increase its affinity for plasmin. A set of computationally designed SAK mutants was produced recombinantly and biochemically characterized. Screening revealed a pharmacologically interesting SAK mutant with-7-fold enhanced affinity toward plasmin,-10-fold improved plasmin selectivity and moderately higher plasmin-generating efficiency in vitro. Collectively, the results obtained provide a framework for SAK engineering using computational affinity-design that could pave the way to next-generation of effective, highly selective, and less toxic thrombolytics.
ER  -

NIKITIN, Dmitri; Jan MIČAN; Martin TOUL; David BEDNÁŘ; Michaela PEŠKOVÁ; Patrícia KITTOVÁ; Sandra THALEROVÁ; Jan VÍTEČEK; Jiří DAMBORSKÝ; Robert MIKULÍK; Sarel J. FLEISHMAN; Zbyněk PROKOP a Martin MAREK. Computer-aided engineering of staphylokinase toward enhanced affinity and selectivity for plasmin. \textit{Computational and Structural Biotechnology Journal}. Amsterdam: Elsevier, 2022, roč.~20, May 2022, s.~1366-1377. ISSN~2001-0370. Dostupné z: https://doi.org/10.1016/j.csbj.2022.03.004.

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