Importance of Residue 13 and the C-Terminus for the Structure and Activity of the Antimicrobial Peptide Aurein 2.2

John T.J. Cheng, John D. Hale, Jason Kindrachuk, Håvard Jenssen, Melissa Elliott, Robert E.W. Hancock, Suzana K. Straus

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

    Resumé

    Previous studies on aurein 2.2 and 2.3 in DMPC/DMPG and POPC/POPG membranes have shown that bilayer thickness and phosphatidylglycerol content have a significant impact on the interaction of these peptides with membrane bilayers. Further examination with the DiSC35 assay has indicated that aurein 2.2 induces greater membrane leakage than aurein 2.3 in Staphylococcus aureus C622. The only difference between these peptides is a Leu to Ile mutation at residue 13. To better understand the importance of this residue, the structure and activity of the L13A, L13F, and L13V mutants were investigated. In addition, we investigated a number of peptides with truncations at the C-terminus to determine whether the C-terminus, which contains residue 13, is crucial for antimicrobial activity. Solution circular dichroism results demonstrated that the L13F mutation and the truncation of the C-terminus by six residues resulted in decreased helical content, whereas the L13A or L13V mutation and the truncation of the C-terminus by three residues showed little to no effect on the structure. Oriented circular dichroism results demonstrated that only an extensive C-terminal truncation reduced the ability of the peptide to insert into lipid bilayers. 31P NMR spectroscopy showed that all peptides disorder the headgroups. The implications of these results in terms of antimicrobial
    activity and the ability of these peptides to induce leakage in S. aureus are discussed. The results suggest that the presence of the 13th residue in aurein 2.2 is important for structure and activity, but the exact nature of residue 13 is less important as long as it is a hydrophobic residue.
    OriginalsprogEngelsk
    TidsskriftBiophysical Journal
    Vol/bind99
    Udgave nummer9
    Sider (fra-til)2926-2935
    ISSN0006-3495
    DOI
    StatusUdgivet - 2010

    Citer dette

    Cheng, John T.J. ; Hale, John D. ; Kindrachuk, Jason ; Jenssen, Håvard ; Elliott, Melissa ; Hancock, Robert E.W. ; Straus, Suzana K. . / Importance of Residue 13 and the C-Terminus for the Structure and Activity of the Antimicrobial Peptide Aurein 2.2. I: Biophysical Journal. 2010 ; Bind 99, Nr. 9. s. 2926-2935.
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    title = "Importance of Residue 13 and the C-Terminus for the Structure and Activity of the Antimicrobial Peptide Aurein 2.2",
    abstract = "Previous studies on aurein 2.2 and 2.3 in DMPC/DMPG and POPC/POPG membranes have shown that bilayer thickness and phosphatidylglycerol content have a significant impact on the interaction of these peptides with membrane bilayers. Further examination with the DiSC35 assay has indicated that aurein 2.2 induces greater membrane leakage than aurein 2.3 in Staphylococcus aureus C622. The only difference between these peptides is a Leu to Ile mutation at residue 13. To better understand the importance of this residue, the structure and activity of the L13A, L13F, and L13V mutants were investigated. In addition, we investigated a number of peptides with truncations at the C-terminus to determine whether the C-terminus, which contains residue 13, is crucial for antimicrobial activity. Solution circular dichroism results demonstrated that the L13F mutation and the truncation of the C-terminus by six residues resulted in decreased helical content, whereas the L13A or L13V mutation and the truncation of the C-terminus by three residues showed little to no effect on the structure. Oriented circular dichroism results demonstrated that only an extensive C-terminal truncation reduced the ability of the peptide to insert into lipid bilayers. 31P NMR spectroscopy showed that all peptides disorder the headgroups. The implications of these results in terms of antimicrobial activity and the ability of these peptides to induce leakage in S. aureus are discussed. The results suggest that the presence of the 13th residue in aurein 2.2 is important for structure and activity, but the exact nature of residue 13 is less important as long as it is a hydrophobic residue.",
    author = "Cheng, {John T.J.} and Hale, {John D.} and Jason Kindrachuk and H{\aa}vard Jenssen and Melissa Elliott and Hancock, {Robert E.W.} and Straus, {Suzana K.}",
    year = "2010",
    doi = "10.1016/j.bpj.2010.08.077",
    language = "English",
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    journal = "Biophysical Journal",
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    Importance of Residue 13 and the C-Terminus for the Structure and Activity of the Antimicrobial Peptide Aurein 2.2. / Cheng, John T.J.; Hale, John D.; Kindrachuk, Jason ; Jenssen, Håvard; Elliott, Melissa; Hancock, Robert E.W. ; Straus, Suzana K. .

    I: Biophysical Journal, Bind 99, Nr. 9, 2010, s. 2926-2935.

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

    TY - JOUR

    T1 - Importance of Residue 13 and the C-Terminus for the Structure and Activity of the Antimicrobial Peptide Aurein 2.2

    AU - Cheng, John T.J.

    AU - Hale, John D.

    AU - Kindrachuk, Jason

    AU - Jenssen, Håvard

    AU - Elliott, Melissa

    AU - Hancock, Robert E.W.

    AU - Straus, Suzana K.

    PY - 2010

    Y1 - 2010

    N2 - Previous studies on aurein 2.2 and 2.3 in DMPC/DMPG and POPC/POPG membranes have shown that bilayer thickness and phosphatidylglycerol content have a significant impact on the interaction of these peptides with membrane bilayers. Further examination with the DiSC35 assay has indicated that aurein 2.2 induces greater membrane leakage than aurein 2.3 in Staphylococcus aureus C622. The only difference between these peptides is a Leu to Ile mutation at residue 13. To better understand the importance of this residue, the structure and activity of the L13A, L13F, and L13V mutants were investigated. In addition, we investigated a number of peptides with truncations at the C-terminus to determine whether the C-terminus, which contains residue 13, is crucial for antimicrobial activity. Solution circular dichroism results demonstrated that the L13F mutation and the truncation of the C-terminus by six residues resulted in decreased helical content, whereas the L13A or L13V mutation and the truncation of the C-terminus by three residues showed little to no effect on the structure. Oriented circular dichroism results demonstrated that only an extensive C-terminal truncation reduced the ability of the peptide to insert into lipid bilayers. 31P NMR spectroscopy showed that all peptides disorder the headgroups. The implications of these results in terms of antimicrobial activity and the ability of these peptides to induce leakage in S. aureus are discussed. The results suggest that the presence of the 13th residue in aurein 2.2 is important for structure and activity, but the exact nature of residue 13 is less important as long as it is a hydrophobic residue.

    AB - Previous studies on aurein 2.2 and 2.3 in DMPC/DMPG and POPC/POPG membranes have shown that bilayer thickness and phosphatidylglycerol content have a significant impact on the interaction of these peptides with membrane bilayers. Further examination with the DiSC35 assay has indicated that aurein 2.2 induces greater membrane leakage than aurein 2.3 in Staphylococcus aureus C622. The only difference between these peptides is a Leu to Ile mutation at residue 13. To better understand the importance of this residue, the structure and activity of the L13A, L13F, and L13V mutants were investigated. In addition, we investigated a number of peptides with truncations at the C-terminus to determine whether the C-terminus, which contains residue 13, is crucial for antimicrobial activity. Solution circular dichroism results demonstrated that the L13F mutation and the truncation of the C-terminus by six residues resulted in decreased helical content, whereas the L13A or L13V mutation and the truncation of the C-terminus by three residues showed little to no effect on the structure. Oriented circular dichroism results demonstrated that only an extensive C-terminal truncation reduced the ability of the peptide to insert into lipid bilayers. 31P NMR spectroscopy showed that all peptides disorder the headgroups. The implications of these results in terms of antimicrobial activity and the ability of these peptides to induce leakage in S. aureus are discussed. The results suggest that the presence of the 13th residue in aurein 2.2 is important for structure and activity, but the exact nature of residue 13 is less important as long as it is a hydrophobic residue.

    U2 - 10.1016/j.bpj.2010.08.077

    DO - 10.1016/j.bpj.2010.08.077

    M3 - Journal article

    VL - 99

    SP - 2926

    EP - 2935

    JO - Biophysical Journal

    JF - Biophysical Journal

    SN - 0006-3495

    IS - 9

    ER -