Binding kinetics of grouper nervous necrosis viruses with functionalized antimicrobial peptides by nanomechanical detection.
We report the binding kinetics of fish-infected grouper nervous necrosis viruses (NNV) and selected antimicrobial peptides (AMPs) by nanomechanical detection. AMPs, the vital member in an innate immunity, are promising candidates in the fight against pathogens due to their broad range of antimicrobial activity and low toxicity. Grouper NNV primarily cause mass mortality of many marine cultured fish species, and two selected AMPs in this study were found to inhibit viruses by agglutinating its virions to form aggregates. The binding activity of NNVs with functionalized AMPs onto a sensing microcantilever yielded induced surface stresses, indicating high binding strength of molecular interaction. The binding affinity and kinetic rate constants of molecular recognition events calculated for NNV-AMP(TH1-5) compared to NNV-AMP(cSALF) were found to be 2.1-fold and 4.43-fold, respectively, indicating TH1-5 effectively bind with NNV more than cSALF. Moreover, a microscopic X-ray photoelectron spectroscopy technique was employed for further validation of pre- and post-NNV binding onto peptides-functionalized sensing surface. An increase in the spectrum and intensity of the P 2p and N 1s elements for the post-NNV binding was clearly shown to ensure the existence of phosphate groups and nitrogen-containing ring structures of specific NNV-TH1-5 interaction. Therefore, the microcantilever biosensing technique provides a potential and useful screening of AMPs for affinity to NNVs.
Institute of Applied Mechanics, National Taiwan University, Taipei 106, Taiwan.
Shu
S
Kuan
Shau-Chi
SC
Chi
Yung-Jen
YJ
Cheng
Ta-Jui
TJ
Chia
Long-Sun
LS
Huang
D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000378:metabolism*; D001492:Bass / Q000821:virology*; D015374:Biosensing Techniques / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005393:Fish Diseases / Q000175:diagnosis / Q000821:virology*; D055617:Micro-Electrical-Mechanical Systems / Q000295:instrumentation*; D030381:Nodaviridae / Q000302:isolation & purification* / Q000378:metabolism; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D056189:Viral Tropism; D014773:Virology / Q000295:instrumentation; D014777:Virus Diseases / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology
D023181:Antimicrobial Cationic Peptides