Granulysin, a T cell product, kills bacteria by altering membrane permeability.
Granulysin, a protein located in the acidic granules of human NK cells and cytotoxic T cells, has antimicrobial activity against a broad spectrum of microbial pathogens. A predicted model generated from the nuclear magnetic resonance structure of a related protein, NK lysin, suggested that granulysin contains a four alpha helical bundle motif, with the alpha helices enriched for positively charged amino acids, including arginine and lysine residues. Denaturation of the polypeptide reduced the alpha helical content from 49 to 18% resulted in complete inhibition of antimicrobial activity. Chemical modification of the arginine, but not the lysine, residues also blocked the antimicrobial activity and interfered with the ability of granulysin to adhere to Escherichia coli and Mycobacterium tuberculosis. Granulysin increased the permeability of bacterial membranes, as judged by its ability to allow access of cytosolic ss-galactosidase to its impermeant substrate. By electron microscopy, granulysin triggered fluid accumulation in the periplasm of M. tuberculosis, consistent with osmotic perturbation. These data suggest that the ability of granulysin to kill microbial pathogens is dependent on direct interaction with the microbial cell wall and/or membrane, leading to increased permeability and lysis.
Ernst
W A
WA
Division of Dermatology, Division of Pulmonary Medicine, Department of Microbiology and Immunology, and Molecular Biology Institute, University of California School of Medicine, Los Angeles, CA 90095, USA.
Thoma-Uszynski
S
S
Teitelbaum
R
R
Ko
C
C
Hanson
D A
DA
Clayberger
C
C
Krensky
A M
AM
Leippe
M
M
Bloom
B R
BR
Ganz
T
T
Modlin
R L
RL
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000138:chemical synthesis / Q000633:toxicity*; D000945:Antigens, Differentiation, T-Lymphocyte / Q000096:biosynthesis / Q000235:genetics / Q000633:toxicity*; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000276:immunology*; D004926:Escherichia coli / Q000187:drug effects / Q000254:growth & development*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000254:growth & development* / Q000648:ultrastructure; D009994:Osmolar Concentration; D010455:Peptides / Q000138:chemical synthesis / Q000633:toxicity; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000138:chemical synthesis / Q000302:isolation & purification; D016176:T-Lymphocyte Subsets / Q000276:immunology* / Q000382:microbiology
D000900:Anti-Bacterial Agents; D000945:Antigens, Differentiation, T-Lymphocyte; C075005:GNLY protein, human; D010455:Peptides; D011994:Recombinant Proteins