Title

Synthesis of antimicrobial TAT multimers

Date of Publication

2019

Document Type

Master's Thesis

Degree Name

Master of Science in Chemistry

Subject Categories

Amino Acids, Peptides, and Proteins | Cells | Medicine and Health Sciences

College

College of Science

Department/Unit

Chemistry

Thesis Adviser

Glenn V. Alea
Jonel P. Saludes

Defense Panel Chair

Marissa G. Noel

Defense Panel Member

Vincent Ng
Mafel C. Ysrael
Jaime Raul Janairo

Abstract/Summary

Cell penetrating peptides (CPP) are short cationic peptides that offer therapeutic use as drug cargoes through their innate ability to permeate the cell membrane. The tridecapeptide (GRKKRRQRRRPPQ) is a known sequence from the 86 to 102 amino acid long protein from the human immunodeficiency virus (HIV)-1 TAT at residues 48 to 60. This peptide has been shown to be an effective antibacterial agent (MIC₅₀ at 2-8 Μm) and cellular translocation in the monomer form. Since better bactericidal action and permeation is observed in prior studies of the TAT to tetramer may further enhance the properties of the peptide. Hence, this study aimed to investigate the antimicrobial activity of TAT peptide multimers. Using bis-Fmoc protected lysine allowed for the synthesis of the multimers in a non-linear approach to maximize the efficiency of the synthesis. The monomeric, dimeric, tetrameric TAT peptides were synthesized through solid phase peptide synthesis, purified by reverse phase high performance liquid chromatography and characterized by mass spectrometry. Also, the secondary structure of the multimeric CPPs were investigated using circular dichroism (CD). The far-UV CD spectra of the multimers revealed that all the synthesized CPPs followed a polyproline-II helix. Branched dimerization of TAT using bis-Fmoc protected lysine allowed the TAT units to retain it monomeric identity even upon mutlimerization. The synthesized peptides were also tested for antimicrobial activity against Staphylococcus aureus, Esherichia coli and Staphylococcus saprophyticus using microbroth susceptibility and minimum inhibitory concentration test. All three test pathogens were found to be non-susceptible against the monomeric peptide. However, S. saprophyticus had shown susceptibility with the TAT peptides dimer and tetramer. The MIC₅₀ and MIC₉₀ of the dimer against the susceptible pathogen was 12.50 μM and 6.25 μM, respectively. The tetrameric TAT demonstrated 90% growth inhibition of S. sap

Abstract Format

html

Language

English

Format

Print

Accession Number

TG07335

Shelf Location

Archives, The Learning Commons, 12F Henry Sy Sr. Hall

Physical Description

xiv, 163 leaves ; illustrations (some color) ; 28 cm.

Keywords

Peptides--Synthesis; Anti-infective agents

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