Introduction Introduction What is antimicrobial peptide (1 slide) Peptide HHC-36, hydrophilic and hydrophobic tail. Explain the pic which part is hydrophilic and which part is hydrophobic? (2 slides) K R K W W R R (W) Rewrite the methods, result and discussion part for power point. Methods (3 slides) Membrane mimetic preparation Measured two bacteria (E-coli and POPC: POPG) and two mammalian (POPG: Cholesterol and POPC) difference amount and in four flakes. dissolved in chloroform and methanol. • • Dry rotary evaporator. • • Dissolved in aqueous buffer. Cycle with Sonicate, thaw, and freeze, five times. Preparation of carboxyfluorescein Weighted carboxyfluorescein and added aqueous buffer then. Stirring the solution with a stie bar Dissolved and adjust final PH value. Preparation of the size-exclusion chromatograph column. In the container mixed sephadex G-25 resin and buffer left to hydrated while spinning and shaking gentil all night. Added buffer and the column was to flow slowly and continuously while the aliquot was being introduce. Preparation of liposome with carboxyfluorescein. Measured POPC, dissolved in chloroform and methanol and dried. Added carboxyflurescein solution. Cycle with freeze, thaw, and sonication five times. Used the extruderand filled the injector syringe with the liposome and injected them through the extruder, passing them between the syringes 11 times. In the equilibrate the gravity sizing column gently added the liposome and stopped the column flow. Added buffer in the top of the beads to fill up the column. The dark orange/greenish coloured liposome sample and the resin fraction intact. Liposome disruption assay Setup the fluorescence spectrophotometer recorded emission of fluorescence at 512nm and 30 min. first a cuvette was filled with 95 μl of buffer. Added liposome after few min and Then added peptide. Finally added the triton-x1000. Result (1 slides) Fluorescence spectrometer reading 600 fluoresence intensity 500 400 300 200 100 0 0 100 200 -100 -200 POPC AND POPG 60μ 300 400 500 wavelengh (nm) ECOLI 60 Intensity (a.u.) • POPC 60μ Intensity (a.u.) • POPC AND COLESTROLE 60μl μl Figure 1 illustration the fluorescence spectrometer reading for different liposomes. Liposome disruption WAVELENGH 350 300 250 200 150 100 50 0 POPC PEPTIDE LYSIS ASSAY INTESITY Figure 2 For carboxyfluorescein-loaded vesicles made from two lipid compositions pure POPC and a combination of POPC: POPG 8:2 ratio steady-state fluorescence spectra were acquired. Discussion (2 slides) peptide HHC-36 the interactions with various model lipid membranes through biophysical measurements. From the CD spectroscopy and fluorescence. The experiment result was unsuccessful because. According to the figure 1, Peptide HHC -36 was interact with POPC: CHLOSTROL and POPC. That means our peptide can destroy human being cell. According to liposome disruption assay graph, the results indicated that HHC-36 did significantly disrupt POPC vesicle HHC-36 affected permeabilization and leakage of calcein from the POPC or human cell. This suggests. there is no obvious trend and therefore indicating there were some errors when the experiment was carried out. Conclusion In conclusion, despite the limitations time and even the result of study unsuccessful