Sulphated penta-galloyl glucopyranoside (SPGG) is glycosaminoglycan mimetic allosteric inhibitor of cathepsin G.
Sulphated penta-galloyl glucopyranoside (SPGG) is glycosaminoglycan mimetic allosteric inhibitor of cathepsin G.
复制标题
硫酸化五没食子酰吡喃葡萄糖苷 (SPGG) 是组织蛋白酶 G 的糖胺聚糖模拟变构抑制剂。
DOI：
10.1093/rpsppr/rqad001
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发表时间：
2023-01
期刊：
影响因子：
--
通讯作者：
Mottamal, Madhusoodanan
中科院分区：
文献类型：
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作者：
Al-Horani, Rami A;Afosah, Daniel K;Kar, Srabani;Aliter, Kholoud F;Mottamal, Madhusoodanan
组织蛋白酶G（CatG）是一种具有广泛底物特异性的阳离子丝氨酸蛋白酶。据报道，CatG在几种炎性病理中起作用。因此，我们的目的是确定一种有效的和变构的CatG抑制剂，用作进一步药物开发机会的平台。显色底物水解试验用于评估SPGG对CatG的抑制效力和选择性。利用盐依赖性研究、Michaelis-Menten动力学和SDS-PAGE来解释SPGG抑制CatG的机制。分子建模也被用来确定一个合理的结合位点。SPGG显示出对CatG的57 nM的抑制效力，其相对于其他蛋白酶具有实质性的选择性。SPGG保护纤连蛋白和层粘连蛋白免受CatG介导的降解。SPGG减少VMAX的CatG水解显色底物，而不影响KM，这表明变构机制。能量贡献的分辨率表明，非离子相互作用贡献了约91%的结合能，这表明特异性识别的可能性很大。分子模拟表明，SPGG可与109 SRRVRRNRN 117的阴离子结合序列结合。我们目前发现SPGG作为CatG的第一个小分子、强效、变构糖胺聚糖模拟物抑制剂。SPGG有望开辟临床相关别构CatG抗炎剂的主要途径。
Cathepsin G (CatG) is a cationic serine protease with wide substrate specificity. CatG is reported to play a role in several inflammatory pathologies. Thus, we aimed at identifying a potent and allosteric inhibitor of CatG to be used as a platform in further drug development opportunities. Chromogenic substrate hydrolysis assays were used to evaluate the inhibition potency and selectivity of SPGG towards CatG. Salt-dependent studies, Michaelis–Menten kinetics and SDS-PAGE were exploited to decipher the mechanism of CatG inhibition by SPGG. Molecular modelling was also used to identify a plausible binding site. SPGG displayed an inhibition potency of 57 nM against CatG, which was substantially selective over other proteases. SPGG protected fibronectin and laminin against CatG-mediated degradation. SPGG reduced VMAX of CatG hydrolysis of a chromogenic substrate without affecting KM, suggesting an allosteric mechanism. Resolution of energy contributions indicated that non-ionic interactions contribute ~91% of binding energy, suggesting a substantial possibility of specific recognition. Molecular modelling indicated that SPGG plausibly binds to an anion-binding sequence of 109SRRVRRNRN117. We present the discovery of SPGG as the first small molecule, potent, allosteric glycosaminoglycan mimetic inhibitor of CatG. SPGG is expected to open a major route to clinically relevant allosteric CatG anti-inflammatory agents.