A High-Throughput Small-Angle X-ray Scattering Assay to Determine the Conformational Change of Plasminogen.
SAXS
conformational change
fibrinolysis
kringle domain
lysine analogue
lysine binding site
plasminogen
structure-function
Journal
International journal of molecular sciences
ISSN: 1422-0067
Titre abrégé: Int J Mol Sci
Pays: Switzerland
ID NLM: 101092791
Informations de publication
Date de publication:
19 Sep 2023
19 Sep 2023
Historique:
received:
12
08
2023
revised:
09
09
2023
accepted:
11
09
2023
medline:
29
9
2023
pubmed:
28
9
2023
entrez:
28
9
2023
Statut:
epublish
Résumé
Plasminogen (Plg) is the inactive form of plasmin (Plm) that exists in two major glycoforms, referred to as glycoforms I and II (GI and GII). In the circulation, Plg assumes an activation-resistant "closed" conformation via interdomain interactions and is mediated by the lysine binding site (LBS) on the kringle (KR) domains. These inter-domain interactions can be readily disrupted when Plg binds to lysine/arginine residues on protein targets or free L-lysine and analogues. This causes Plg to convert into an "open" form, which is crucial for activation by host activators. In this study, we investigated how various ligands affect the kinetics of Plg conformational change using small-angle X-ray scattering (SAXS). We began by examining the open and closed conformations of Plg using size-exclusion chromatography (SEC) coupled with SAXS. Next, we developed a high-throughput (HTP) 96-well SAXS assay to study the conformational change of Plg. This method enables us to determine the
Identifiants
pubmed: 37762561
pii: ijms241814258
doi: 10.3390/ijms241814258
pmc: PMC10531915
pii:
doi:
Substances chimiques
Plasminogen
9001-91-6
Ligands
0
Lysine
K3Z4F929H6
Serine Proteases
EC 3.4.-
Antibodies
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : National Health and Medical Research Council
ID : APP1127593, APP1029403
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