NRC-04 Lab Report

ABSTRACT
NRC-04, a novel antimicrobial peptide derived from skin mucous secretions of flat fish winter flounder, shows a broad spectrum of antimicrobial activity. In order to understand the conformational change of NRC-04 in different types of membrane, our team did experiments on NRC-04 with negatively charged bacterial surface membrane mimetic micelles sodium dodecyl sulphate(SDS), zwitterionic eukaryotic middle membrane mimetic micelles dodecylphosphocholine(DPC), gram-negative bacteria outer membrane mimetic micelles Lipopolysaccharide(LPS) and bacterial inner membrane mimetic micelles 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol(POPG). Fluorescence test shows that the C-terminus tryptophan residue of NRC-04 interacts with the hydrophobic

Figure 20 and 21 casting tray Figure 22 gel box
After 1 hr., take out the gel from gel box carefully, place it into the machine, so that the DNA gel electrophoresis can be visualize under UV light. Figure 23 gel documentation system, used to visualize gel electrophoresis with UV light
NMR spectroscopy
First of all, 3 samples were prepared, peptide in SDS, DPC, and buffer. The sample temperature was maintained at 298 K, prepared by supervisor and H(hydrogen) in SDS and DPC micelle was replaced with D(deuterium) , so that in proton NMR, peptide won’t be interfered by H in micelle. amount of peptide in sds and dpc
In the case of NMR of peptide alone, sample was prepared with 90% of H2O which is 540μL and 10% of D2O which is 60μL, so 600μL of solution was used to dilute 1.5mg of peptide.
Put 600μL of sample into NMR Sample Tubes, put the sample tube into NMR sample holder, and then run the test, the chemical shift of proton in peptide can be monitored.
NMR measurements were performed on Bruker Bruker Avance DRX 800 [32]
Figure 24 NMR machine Bruker Bruker Avance DRX