Rational design of transglutaminase substrate peptides for rapid enzymatic formation of hydrogels
Rational design of transglutaminase substrate peptides for rapid enzymatic formation of hydrogels
复制标题
合理设计转谷氨酰胺酶底物肽，用于快速酶解形成水凝胶
DOI：
10.1021/ja038593b
复制
发表时间：
2003-11-26
影响因子：
15
通讯作者：
Messersmith, PB
中科院分区：
文献类型：
--
作者：
Hu, BH;Messersmith, PB
设计、表征了对转氨酶（TGase）具有高特异性的短肽底物，并将其偶联至生物相容性聚合物，从而允许肽-聚合物缀合物快速酶促交联成水凝胶。合理设计并合成了8种酰基受体赖氨酸-肽底物和3种酰基供体谷氨酰胺-肽底物。这些肽对组织转氨酶的动力学常数进行了测定，通过酶测定使用RP-HPLC分析与LC-ESI/MS的帮助下。几个酰基供体和酰基受体肽对TGase具有高特异性被确定，包括一些含有不寻常的氨基酸1 - 3，4-二羟基苯丙氨酸（DOPA），这是发现在海洋和淡水贻贝分泌的粘附蛋白。将具有高底物特异性的酰基供体和酰基受体肽分别偶联到支链聚乙二醇（PEG）聚合物分子上。这些聚合物-肽缀合物的等摩尔溶液在生理条件下在转氨酶存在下在不到2分钟内迅速形成水凝胶。生物相容性结构单元的使用、它们在生理条件下从液体前体的快速固化、以及使用生物良性酶交联掺入粘附性氨基酸残基的能力，对于将此类材料用于组织修复、药物递送和组织工程应用是有利的性质。
Short peptide substrates with high specificity toward transglutaminase (TGase) enzyme were designed, characterized, and coupled to a biocompatible polymer, allowing for rapid enzymatic cross-linking of peptide-polymer conjugates into hydrogels. Eight acyl acceptor Lys-peptide substrates and three acyl donor Gln-peptide substrates were rationally designed and synthesized. The kinetic constants of these peptides toward tissue transglutaminase were measured by enzyme assay using RP-HPLC analysis with the aid of LC-ESI/MS. Several acyl donor and acyl acceptor peptides with high specificities toward TGase were identified, including a few containing the unusual amino acidl-3,4-dihydroxylphenylalanine (DOPA), which is found in the adhesive proteins secreted by marine and freshwater mussels. Acyl donor and acyl acceptor peptides with high substrate specificities were separately coupled to branched poly(ethylene glycol) (PEG) polymer molecules. Equimolar solutions of these polymer-peptide conjugates rapidly formed hydrogels in less than 2 min in the presence of transglutaminase under physiological conditions. The use of biocompatible building blocks, their rapid solidification from a liquid precursor under physiologic conditions, and the ability to incorporate adhesive amino acid residues using biologically benign enzymatic cross-linking are advantageous properties for the use of such materials for tissue repair, drug delivery, and tissue engineering applications.