Assessing the impact of His-tags on activity and stability of staphylokinase variants
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00159816%3A_____%2F25%3A00082379" target="_blank" >RIV/00159816:_____/25:00082379 - isvavai.cz</a>
Alternative codes found
RIV/00216224:14310/25:00142671
Result on the web
<a href="https://www.sciencedirect.com/science/article/pii/S0141813025082121" target="_blank" >https://www.sciencedirect.com/science/article/pii/S0141813025082121</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/j.ijbiomac.2025.147655" target="_blank" >10.1016/j.ijbiomac.2025.147655</a>
Alternative languages
Result language
angličtina
Original language name
Assessing the impact of His-tags on activity and stability of staphylokinase variants
Original language description
Staphylokinase (SAK), a potent plasminogen activator, is a promising thrombolytic agent, but its clinical application is limited by immunogenicity and stability concerns. In addition to intrinsic sequence variants, recombinant protein production often introduces affinity tags, such as the N-terminal polyhistidine (His-tag), whose potential effects on protein's biophysical and functional properties remain poorly understood. Here, we systematically investigated the impact of His-tagging on the stability and activity of four SAK variants: wild-type and non-immunogenic (triple-alanine, 3A) forms of two naturally occurring SAK types, SAK STAR and SAK 42D. Thermal and pH stability were assessed using circular dichroism and tryptophan fluorescence spectroscopy, and plasminogen-activating efficiency was evaluated through chromogenic assays. We found that while the His-tag had little effect on thermal stability and only modestly influenced functional activity, it significantly destabilized SAK under acidic conditions, and altered unfolding transitions, indicating the presence of intermediate conformations. Among the tested proteins, SAK STAR demonstrated the best structural and functional robustness, whereas SAK 42D 3A was the least stable and most prone to aggregation. These results highlight the need to assess the biophysical effect of affinity tags and point SAK STAR as the most suitable candidate for next therapeutic development.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10608 - Biochemistry and molecular biology
Result continuities
Project
<a href="/en/project/LX22NPO5107" target="_blank" >LX22NPO5107: National institute for Neurological Research</a><br>
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2025
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
International Journal of Biological Macromolecules
ISSN
0141-8130
e-ISSN
1879-0003
Volume of the periodical
328
Issue of the periodical within the volume
Nov 2025
Country of publishing house
NL - THE KINGDOM OF THE NETHERLANDS
Number of pages
10
Pages from-to
147655
UT code for WoS article
001576696500001
EID of the result in the Scopus database
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