Surface Plasmon Resonance for the Interaction of Capsular Polysaccharide (CPS) With KpACE

The study of carbohydrate–protein interactions is crucial for clarifying biological processes and identifying potential drug candidates. However, due to the complex structure of carbohydrates, such as high molecular weight, dynamic flexibility, and high solution viscosity, it is challenging to study...

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Main Authors: Zhe Wang, Lijun Wang, Xiaomin Zhang, Jing- Zhang, Chao Cai
Format: Article
Language:English
Published: Bio-protocol LLC 2025-06-01
Series:Bio-Protocol
Online Access:https://bio-protocol.org/en/bpdetail?id=5346&type=0
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author Zhe Wang
Lijun Wang
Xiaomin Zhang
Jing- Zhang
Chao Cai
author_facet Zhe Wang
Lijun Wang
Xiaomin Zhang
Jing- Zhang
Chao Cai
author_sort Zhe Wang
collection DOAJ
description The study of carbohydrate–protein interactions is crucial for clarifying biological processes and identifying potential drug candidates. However, due to the complex structure of carbohydrates, such as high molecular weight, dynamic flexibility, and high solution viscosity, it is challenging to study their interactions with diverse proteins. Conventional analytical techniques like isothermal titration calorimetry (ITC), X-ray crystallography, molecular dynamics (MD) simulations, and nuclear magnetic resonance (NMR) spectroscopy have limitations in revealing these molecular interactions. Surface plasmon resonance (SPR), an advanced optical biosensor technique, overcomes these limitations. It enables real-time, label-free monitoring of the interaction dynamics between carbohydrates and proteins through a continuous flow over a chip surface. In this study, we utilized SPR-based techniques to explore the interaction of capsular polysaccharides (CPS) of Klebsiella pneumoniae and the enzyme KpACE (K. pneumoniae acetylated capsule esterase). Our SPR-based analytical platform has several advantages, including shorter experimental time, a simulated physiological state, and minimal sample requirements for investigating carbohydrate–protein interactions. This approach expands the applicability scope of SPR technology and provides a valuable tool for a wide range of research. By using SPR, we successfully verified that KpACE acts on the acetyl groups of CPS, demonstrating its enzymatic activity, which is crucial for understanding the pathogenic mechanism of K. pneumoniae and developing potential antibacterial drugs.
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spelling doaj-art-142cb2eb488c4c74a58e624096e1570d2025-08-20T02:20:52ZengBio-protocol LLCBio-Protocol2331-83252025-06-01151210.21769/BioProtoc.5346Surface Plasmon Resonance for the Interaction of Capsular Polysaccharide (CPS) With KpACEZhe Wang0Lijun Wang1Xiaomin Zhang2Jing- Zhang3Chao Cai4Key Laboratory of Marine Drugs of Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Qingdao, ChinaShandong Key Laboratory of Glycoscience and Glycotherapeutics, School of Medicine and Pharmacy, Ocean University of China, Qingdao, ChinaDepartment of Laboratory Medicine, Beijing Xiaotangshan Hospital, Beijing, ChinaCenter for Infection Biology, School of Basic Medical Sciences, Tsinghua Medicine, Tsinghua University, Beijing, ChinaKey Laboratory of Marine Drugs of Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Qingdao, ChinaCenter for Infection Biology, School of Basic Medical Sciences, Tsinghua Medicine, Tsinghua University, Beijing, ChinaKey Laboratory of Marine Drugs of Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Qingdao, ChinaShandong Key Laboratory of Glycoscience and Glycotherapeutics, School of Medicine and Pharmacy, Ocean University of China, Qingdao, ChinaThe study of carbohydrate–protein interactions is crucial for clarifying biological processes and identifying potential drug candidates. However, due to the complex structure of carbohydrates, such as high molecular weight, dynamic flexibility, and high solution viscosity, it is challenging to study their interactions with diverse proteins. Conventional analytical techniques like isothermal titration calorimetry (ITC), X-ray crystallography, molecular dynamics (MD) simulations, and nuclear magnetic resonance (NMR) spectroscopy have limitations in revealing these molecular interactions. Surface plasmon resonance (SPR), an advanced optical biosensor technique, overcomes these limitations. It enables real-time, label-free monitoring of the interaction dynamics between carbohydrates and proteins through a continuous flow over a chip surface. In this study, we utilized SPR-based techniques to explore the interaction of capsular polysaccharides (CPS) of Klebsiella pneumoniae and the enzyme KpACE (K. pneumoniae acetylated capsule esterase). Our SPR-based analytical platform has several advantages, including shorter experimental time, a simulated physiological state, and minimal sample requirements for investigating carbohydrate–protein interactions. This approach expands the applicability scope of SPR technology and provides a valuable tool for a wide range of research. By using SPR, we successfully verified that KpACE acts on the acetyl groups of CPS, demonstrating its enzymatic activity, which is crucial for understanding the pathogenic mechanism of K. pneumoniae and developing potential antibacterial drugs.https://bio-protocol.org/en/bpdetail?id=5346&type=0
spellingShingle Zhe Wang
Lijun Wang
Xiaomin Zhang
Jing- Zhang
Chao Cai
Surface Plasmon Resonance for the Interaction of Capsular Polysaccharide (CPS) With KpACE
Bio-Protocol
title Surface Plasmon Resonance for the Interaction of Capsular Polysaccharide (CPS) With KpACE
title_full Surface Plasmon Resonance for the Interaction of Capsular Polysaccharide (CPS) With KpACE
title_fullStr Surface Plasmon Resonance for the Interaction of Capsular Polysaccharide (CPS) With KpACE
title_full_unstemmed Surface Plasmon Resonance for the Interaction of Capsular Polysaccharide (CPS) With KpACE
title_short Surface Plasmon Resonance for the Interaction of Capsular Polysaccharide (CPS) With KpACE
title_sort surface plasmon resonance for the interaction of capsular polysaccharide cps with kpace
url https://bio-protocol.org/en/bpdetail?id=5346&type=0
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