Institute for Organic Chemistry and Biochemistry, Technical University of Darmstadt, Alarich-Weiss-Str. 4, 64287 Darmstadt, Germany.
Institute for Macromolecular and Paper Chemistry, Technical University of Darmstadt, Alarich-Weiss-Str. 8, 64287 Darmstadt, Germany.
Biomacromolecules. 2021 Jul 12;22(7):2954-2962. doi: 10.1021/acs.biomac.1c00354. Epub 2021 Jun 8.
Herein, we report a novel two-step method for the covalent, site-directed, and efficient immobilization of proteins on lab-made paper sheets. First, paper fibers were modified with a peptidic anchor comprising enzyme recognition motifs. Four different conjugation strategies for peptide immobilization were evaluated with respect to reproducibility and fiber loading efficiency. After manufacturing of the peptide-preconditioned paper, oriented conjugation of the model protein tGFP containing a C-terminal recognition sequence for either sortase A or microbial transglutaminase was assessed semiquantitatively by fluorescence measurement and inspected by confocal laser scanning microscopy (CLSM). The two enzymes utilized for protein conjugation used the same oligoglycine peptide anchor, and both proved to be suitable for controlled oriented linkage of substrate proteins at physiological conditions.
在此，我们报告了一种新颖的两步法，用于将蛋白质共价、定点、高效地固定在实验室制造的纸页上。首先，用包含酶识别基序的肽锚定修饰纸纤维。针对肽固定的重现性和纤维负载效率，评估了四种不同的偶联策略。在肽预处理纸制造之后，通过荧光测量和共聚焦激光扫描显微镜 (CLSM) 检查，半定量评估了含有针对溶菌酶或微生物转谷氨酰胺酶的 C 末端识别序列的模型蛋白 tGFP 的定向偶联。用于蛋白质偶联的两种酶都使用相同的寡甘氨酸肽锚定，并且都被证明适合在生理条件下控制底物蛋白的定向连接。
