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Title: The elastase-PK101 structure: Mechanism of an ultrasensitive activity-based probe revealed

Abstract

Human neutrophil elastase (HNE) plays a central role in neutrophil host defense, but its broad specificity makes HNE a difficult target for both inhibitor and probe development. Recently, we identified the unnatural amino acid containing activity-based probe PK101, which exhibits astounding sensitivity and selectivity for HNE, yet completely lacks mechanistic explanation for its unique characteristics. Here, we present the crystal structure of the HNE-PK101 complex which not only reveals the basis for PK101 ultrasensitivity but also uncovers so far unrecognized HNE features. Strikingly, the Nle( O-Bzl) function in the P4 position of PK101 reveals and leverages an “exo-pocket” on HNE as a critical factor for selectivity. Furthermore, the PK101 P3 position harbors a methionine dioxide function, which mimics a post-translationally oxidized methionine residue and forms a critical hydrogen bond to the backbone amide of Gly219 of HNE. Gly219 resides in a Gly–Gly motif that is unique to HNE, yet compulsory for this interaction. Consequently, this feature enables HNE to accommodate substrates that have undergone methionine oxidation, which constitutes a hallmark post-translational modification of neutrophil signaling.

Authors:
 [1];  [2];  [3];  [3];  [1]
  1. Sanford-Burnham Medical Research Institute, La Jolla, CA (United States)
  2. Brookhaven National Lab. (BNL), Upton, NY (United States)
  3. Wroclaw Univ. of Technology, Wroclaw (Poland)
Publication Date:
Research Org.:
Brookhaven National Laboratory (BNL), Upton, NY (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1193225
Report Number(s):
BNL-108110-2015-JA
Journal ID: ISSN 1554-8929; R&D Project: LS001
Grant/Contract Number:  
SC00112704
Resource Type:
Accepted Manuscript
Journal Name:
ACS Chemical Biology
Additional Journal Information:
Journal Volume: 10; Journal Issue: 4; Journal ID: ISSN 1554-8929
Publisher:
American Chemical Society
Country of Publication:
United States
Language:
English
Subject:
36 MATERIALS SCIENCE

Citation Formats

Lechtenberg, Bernhard C., Robinson, Howard R., Kasperkiewicz, Paulina, Drag, Marcin, and Riedl, Stefan J. The elastase-PK101 structure: Mechanism of an ultrasensitive activity-based probe revealed. United States: N. p., 2015. Web. doi:10.1021/cb500909n.
Lechtenberg, Bernhard C., Robinson, Howard R., Kasperkiewicz, Paulina, Drag, Marcin, & Riedl, Stefan J. The elastase-PK101 structure: Mechanism of an ultrasensitive activity-based probe revealed. United States. doi:10.1021/cb500909n.
Lechtenberg, Bernhard C., Robinson, Howard R., Kasperkiewicz, Paulina, Drag, Marcin, and Riedl, Stefan J. Thu . "The elastase-PK101 structure: Mechanism of an ultrasensitive activity-based probe revealed". United States. doi:10.1021/cb500909n. https://www.osti.gov/servlets/purl/1193225.
@article{osti_1193225,
title = {The elastase-PK101 structure: Mechanism of an ultrasensitive activity-based probe revealed},
author = {Lechtenberg, Bernhard C. and Robinson, Howard R. and Kasperkiewicz, Paulina and Drag, Marcin and Riedl, Stefan J.},
abstractNote = {Human neutrophil elastase (HNE) plays a central role in neutrophil host defense, but its broad specificity makes HNE a difficult target for both inhibitor and probe development. Recently, we identified the unnatural amino acid containing activity-based probe PK101, which exhibits astounding sensitivity and selectivity for HNE, yet completely lacks mechanistic explanation for its unique characteristics. Here, we present the crystal structure of the HNE-PK101 complex which not only reveals the basis for PK101 ultrasensitivity but also uncovers so far unrecognized HNE features. Strikingly, the Nle(O-Bzl) function in the P4 position of PK101 reveals and leverages an “exo-pocket” on HNE as a critical factor for selectivity. Furthermore, the PK101 P3 position harbors a methionine dioxide function, which mimics a post-translationally oxidized methionine residue and forms a critical hydrogen bond to the backbone amide of Gly219 of HNE. Gly219 resides in a Gly–Gly motif that is unique to HNE, yet compulsory for this interaction. Consequently, this feature enables HNE to accommodate substrates that have undergone methionine oxidation, which constitutes a hallmark post-translational modification of neutrophil signaling.},
doi = {10.1021/cb500909n},
journal = {ACS Chemical Biology},
number = 4,
volume = 10,
place = {United States},
year = {2015},
month = {1}
}

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