Megia-Fernandez Alicia, Mills Bethany, Michels Chesney, Chankeshwara Sunay V, Dhaliwal Kevin, Bradley Mark
EaStChem, School of Chemistry, University of Edinburgh, Joseph Black Building, David Brewster Road, Edinburgh, EH9 3FJ, UK.
Org Biomol Chem. 2017 May 23;15(20):4344-4350. doi: 10.1039/c7ob00663b.
A library of FRET-based peptides were prepared and studied as Thrombin substrates. This identified probes that showed selective activation by Thrombin, low fluorescent background signals, stability to Factor Xa, matrix metalloproteases, and primary human inflammatory cell lysates and supernatant. These were selected for further optimization, creating a second generation of fluorogenic probes with improved solubility and Plasmin resistance. The optimised probe allowed the detection of Thrombin activity in ex vivo fibrotic human tissue.
制备了基于荧光共振能量转移（FRET）的肽库，并将其作为凝血酶底物进行研究。这确定了一些探针，这些探针显示出被凝血酶选择性激活、低荧光背景信号、对因子Xa、基质金属蛋白酶以及原代人炎症细胞裂解物和上清液具有稳定性。选择这些探针进行进一步优化，从而创建了第二代具有改善溶解性和抗纤溶酶性能的荧光探针。优化后的探针能够检测人纤维化离体组织中的凝血酶活性。
