Structural and binding studies of SAP-1 protein with heparin.
SAP-1 is a low molecular weight cysteine protease inhibitor (CPI) which belongs to type-2 cystatins family. SAP-1 protein purified from human seminal plasma (HuSP) has been shown to inhibit cysteine and serine proteases and exhibit interesting biological properties, including high temperature and pH stability. Heparin is a naturally occurring glycosaminoglycan (with varied chain length) which interacts with a number of proteins and regulates multiple steps in different biological processes. As an anticoagulant, heparin enhances inhibition of thrombin by the serpin antithrombin III. Therefore, we have employed surface plasmon resonance (SPR) to improve our understanding of the binding interaction between heparin and SAP-1 (protease inhibitor). SPR data suggest that SAP-1 binds to heparin with a significant affinity (KD = 158 nm). SPR solution competition studies using heparin oligosaccharides showed that the binding of SAP-1 to heparin is dependent on chain length. Large oligosaccharides show strong binding affinity for SAP-1. Further to get insight into the structural aspect of interactions between SAP-1 and heparin, we used modelled structure of the SAP-1 and docked with heparin and heparin-derived polysaccharides. The results suggest that a positively charged residue lysine plays important role in these interactions. Such information should improve our understanding of how heparin, present in the reproductive tract, regulates cystatins activity.
|Keywords||Binding affinity; ; Cystatins; ; Heparin; ; Key residues; ; Molecular docking; ; Surface plasmon resonance|
|Journal||Chemical biology & drug design|
|Journal citation||Vol 85 (Issue 3), pp. 404 - 410|
|Publisher||John Wiley & Sons Ltd|
|Digital Object Identifier (DOI)||https://doi.org/10.1111/cbdd.12420|
|Web address (URL)||https://doi.org/10.1111/cbdd.12420|
|Online||21 Aug 2014|
|Online||22 Sep 2014|
|Online||12 Feb 2015|
|Publication process dates|
|Accepted||13 Aug 2014|
|Deposited||15 Jun 2023|
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