一种新型噬菌体编码的溶菌酶KTN6 Gp46的结构与生化特性
Structural and Biochemical Characterization of a New Phage-Encoded Muramidase, KTN6 Gp46.
作者信息
Sanz-Gaitero Marta, De Maesschalck Vincent, Patel Ankur, Longin Hannelore, Van Noort Vera, Rodriguez-Rubio Lorena, van Ryne Michael, Danis-Wlodarczyk Katarzyna, Drulis-Kawa Zuzanna, Mesnage Stephane, van Raaij Mark, Lavigne Rob
机构信息
Centro Nacional de Biotecnologia, Consejo Superior de Investigaciones Cientificas, Madrid, Spain.
Department of Biological Sciences, Munster Technological University, Cork, Ireland.
出版信息
Phage (New Rochelle). 2024 Jun 21;5(2):53-62. doi: 10.1089/phage.2023.0040. eCollection 2024 Jun.
BACKGROUND
Endolysins are phage-encoded lytic enzymes that degrade bacterial peptidoglycan at the end of phage lytic cycles to release new phage particles. These enzymes are being explored as an alternative to small-molecule antibiotics.
METHODS
The crystal structure of KTN6 Gp46 was determined and compared with a ColabFold model. Cleavage specificity was examined using a peptidoglycan digest and reversed-phase high-performance liquid chromatography coupled to mass spectrometry (HPLC/MS).
RESULTS
The structure of KTN6 Gp46 could be determined at 1.4 Å resolution, and key differences in loops of the putative peptidoglycan binding domain were identified in comparison with its closest known homologue, the endolysin of phage SPN1S. Reversed-phase HPLC/MS analysis of the reaction products following peptidoglycan digestion confirmed the muramidase activity of Gp46, consistent with structural predictions.
CONCLUSION
These insights into the structure and function of endolysins further expand the toolbox for endolysin engineering and explore their potential in enzyme-based antibacterial design strategies.
背景
内溶素是噬菌体编码的裂解酶，在噬菌体裂解周期结束时降解细菌肽聚糖以释放新的噬菌体颗粒。这些酶正被探索作为小分子抗生素的替代品。
方法
测定了KTN6 Gp46的晶体结构，并与ColabFold模型进行了比较。使用肽聚糖消化和反相高效液相色谱-质谱联用（HPLC/MS）检测裂解特异性。
结果
KTN6 Gp46的结构可以在1.4 Å分辨率下确定，与其最接近的已知同源物噬菌体SPN1S的内溶素相比，在假定的肽聚糖结合结构域的环中发现了关键差异。肽聚糖消化后反应产物的反相HPLC/MS分析证实了Gp46的溶菌酶活性，与结构预测一致。
结论
这些对内溶素结构和功能的见解进一步扩展了内溶素工程的工具箱，并探索了它们在基于酶的抗菌设计策略中的潜力。
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