Research output: Contribution to journal › Article › Academic › peer-review
Native thrombocidin-1 and unfolded thrombocidin-1 exert antimicrobial activity via distinct structural elements. / Kwakman, Paulus H. S.; Krijgsveld, Jeroen; de Boer, Leonie et al.
In: Journal of biological chemistry, Vol. 286, No. 50, 2011, p. 43506-43514.Research output: Contribution to journal › Article › Academic › peer-review
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TY - JOUR
T1 - Native thrombocidin-1 and unfolded thrombocidin-1 exert antimicrobial activity via distinct structural elements
AU - Kwakman, Paulus H. S.
AU - Krijgsveld, Jeroen
AU - de Boer, Leonie
AU - Nguyen, Leonard T.
AU - Boszhard, Laura
AU - Vreede, Jocelyne
AU - Dekker, Henk L.
AU - Speijer, Dave
AU - Drijfhout, Jan W.
AU - te Velde, Anje A.
AU - Crielaard, Wim
AU - Vogel, Hans J.
AU - Vandenbroucke-Grauls, Christina M. J. E.
AU - Zaat, Sebastian A. J.
PY - 2011
Y1 - 2011
N2 - Chemokines (chemotactic cytokines) can have direct antimicrobial activity, which is apparently related to the presence of a distinct positively charged patch on the surface. However, chemokines can retain antimicrobial activity upon linearization despite the loss of their positive patch, thus questioning the importance of this patch for activity. Thrombocidin-1 (TC-1) is a microbicidal protein isolated from human blood platelets. TC-1 only differs from the chemokine NAP-2/CXCL7 by a two-amino acid C-terminal deletion, but this truncation is crucial for antimicrobial activity. We assessed the structure-activity relationship for antimicrobial activity of TC-1. Reduction of the charge of the TC-1-positive patch by replacing lysine 17 with alanine reduced the activity against bacteria and almost abolished activity against the yeast Candida albicans. Conversely, augmentation of the positive patch by increasing charge density or size resulted in a 2-3-fold increased activity against Staphylococcus aureus, Escherichia coli, and Bacillus subtilis but did not substantially affect activity against C. albicans. Reduction of TC-1 resulted in loss of the folded conformation, but this disruption of the positive patch did not affect antimicrobial activity. Using overlapping 15-mer synthetic peptides, we demonstrate peptides corresponding to the N-terminal part of TC-1 to have similar antimicrobial activity as intact TC-1. Although we demonstrate that the positive patch is essential for activity of folded TC-1, unfolded TC-1 retained antimicrobial activity despite the absence of a positive patch. This activity is probably exerted by a linear peptide stretch in the N-terminal part of the molecule. We conclude that intact TC-1 and unfolded TC-1 exert antimicrobial activity via distinct structural elements
AB - Chemokines (chemotactic cytokines) can have direct antimicrobial activity, which is apparently related to the presence of a distinct positively charged patch on the surface. However, chemokines can retain antimicrobial activity upon linearization despite the loss of their positive patch, thus questioning the importance of this patch for activity. Thrombocidin-1 (TC-1) is a microbicidal protein isolated from human blood platelets. TC-1 only differs from the chemokine NAP-2/CXCL7 by a two-amino acid C-terminal deletion, but this truncation is crucial for antimicrobial activity. We assessed the structure-activity relationship for antimicrobial activity of TC-1. Reduction of the charge of the TC-1-positive patch by replacing lysine 17 with alanine reduced the activity against bacteria and almost abolished activity against the yeast Candida albicans. Conversely, augmentation of the positive patch by increasing charge density or size resulted in a 2-3-fold increased activity against Staphylococcus aureus, Escherichia coli, and Bacillus subtilis but did not substantially affect activity against C. albicans. Reduction of TC-1 resulted in loss of the folded conformation, but this disruption of the positive patch did not affect antimicrobial activity. Using overlapping 15-mer synthetic peptides, we demonstrate peptides corresponding to the N-terminal part of TC-1 to have similar antimicrobial activity as intact TC-1. Although we demonstrate that the positive patch is essential for activity of folded TC-1, unfolded TC-1 retained antimicrobial activity despite the absence of a positive patch. This activity is probably exerted by a linear peptide stretch in the N-terminal part of the molecule. We conclude that intact TC-1 and unfolded TC-1 exert antimicrobial activity via distinct structural elements
U2 - 10.1074/jbc.M111.248641
DO - 10.1074/jbc.M111.248641
M3 - Article
C2 - 22025617
VL - 286
SP - 43506
EP - 43514
JO - Journal of biological chemistry
JF - Journal of biological chemistry
SN - 0021-9258
IS - 50
ER -
ID: 1508589