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Phosphoramidon inhibits the integral membrane protein zinc metalloprotease ZMPSTE24

Journal Article · · Acta Crystallographica. Section D. Structural Biology

The integral membrane protein zinc metalloprotease ZMPSTE24 possesses a completely novel structure, comprising seven long kinked transmembrane helices that encircle a voluminous 14 000 Å3cavity within the membrane. Functionally conserved soluble zinc metalloprotease residues are contained within this cavity. As part of an effort to understand the structural and functional relationships between ZMPSTE24 and soluble zinc metalloproteases, the inhibition of ZMPSTE24 by phosphoramidon [N-(α-rhamnopyranosyl-oxyhydroxyphosphinyl)-Leu-Trp], a transition-state analog and competitive inhibitor of multiple soluble zinc metalloproteases, especially gluzincins, has been characterized functionally and structurally. The functional results, the determination of preliminary IC50values by the use of an intramolecular quenched-fluorescence fluorogenic peptide assay, indicate that phosphoramidon inhibits ZMPSTE24 in a manner consistent with competitive inhibition. The structural results, a 3.85 Å resolution X-ray crystal structure of a ZMPSTE24–phosphoramidon complex, indicate that the overall binding mode observed between phosphoramidon and soluble gluzincins is conserved. Based on the structural data, a significantly lower potency than that observed for soluble gluzincins such as thermolysin and neprilysin is predicted. These results strongly suggest a close relationship between soluble gluzincins and the integral membrane protein zinc metalloprotease ZMPSTE24.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Organization:
NCINIH
OSTI ID:
1463719
Journal Information:
Acta Crystallographica. Section D. Structural Biology, Vol. 74, Issue 8; ISSN 2059-7983
Publisher:
IUCr
Country of Publication:
United States
Language:
ENGLISH

References (41)

Modulation of Ras and a-Factor Function by Carboxyl-Terminal Proteolysis March 1997
On the size of the active site in proteases. I. Papain April 1967
Human CaaX protease ZMPSTE24 expressed in yeast: Structure and inhibition by HIV protease inhibitors: ZMPSTE24 Structure and Inhibition November 2016
Ste24p Mediates Proteolysis of Both Isoprenylated and Non-prenylated Oligopeptides April 2016
HIV protease inhibitors block the zinc metalloproteinase ZMPSTE24 and lead to an accumulation of prelamin A in cells July 2007
Inhibitory effects of phosphoramidon on neutral metalloendopeptidases and its application on affinity chromatography July 1975
Crystallographic structural analysis of phosphoramidates as inhibitors and transition-state analogs of thermolysin June 1986
Features and development of Coot March 2010
Assessing and maximizing data quality in macromolecular crystallography October 2015
Yeast Genes Controlling Responses to Topogenic Signals in a Model Transmembrane Protein April 2002
The Protease Ste24 Clears Clogged Translocons January 2016
Studies on inhibitory effect of phosphoramidon and its analogs on thermolysin December 1975
A Novel Membrane-associated Metalloprotease, Ste24p, Is Required for the First Step of NH 2 -terminal Processing of the Yeast a-Factor Precursor January 1997
Human Ras-Converting Enzyme (hRCE1) Endoproteolytic Activity on K-Ras-Derived Peptides November 2000
Towards automated crystallographic structure refinement with phenix.refine March 2012
Cell-specific activity of neprilysin 2 isoforms and enzymic specificity compared with neprilysin April 2002
PHENIX: a comprehensive Python-based system for macromolecular structure solution January 2010
The lamin protein family January 2011
How good are my data and what is the resolution? June 2013
On the active site of proteases. III. Mapping the active site of papain; specific peptide inhibitors of papain September 1968
Purification of transmembrane proteins from Saccharomyces cerevisiae for X-ray crystallography June 2010
Acquisition of accurate data from intramolecular quenched fluorescence protease assays April 2017
Comprehensive Characterization of Genes Required for Protein Folding in the Endoplasmic Reticulum March 2009
Human ZMPSTE24 disease mutations: residual proteolytic activity correlates with disease severity June 2012
Small-Molecule Inhibitors of the Rce1p CaaX Protease October 2007
Nuclear lamins and laminopathies November 2011
Structure of the Integral Membrane Protein CAAX Protease Ste24p March 2013
A Novel Role of the Yeast CaaX Protease Ste24 in Chitin Synthesis July 2010
Structure of human neutral endopeptidase (neprilysin) complexed with phosphoramidon 1 1Edited by R. Huber February 2000
Prelamin A Acts to Accelerate Smooth Muscle Cell Senescence and Is a Novel Biomarker of Human Vascular Aging May 2010
XDS January 2010
Proteolytic Systems: Constructing Degradomes January 2009
The Structural Basis of ZMPSTE24-Dependent Laminopathies March 2013
A Thermolysin Inhibitor Produced by Actinomycetes: Phosphoramidon January 1973
A Novel Family of Soluble Minimal Scaffolds Provides Structural Insight into the Catalytic Domains of Integral Membrane Metallopeptidases June 2013
Analysis of Prelamin A Biogenesis Reveals the Nucleus to be a CaaX Processing Compartment December 2008
HIV protease inhibitors inhibit FACE1/ZMPSTE24: a mechanism for acquired lipodystrophy in patients on highly active antiretroviral therapy? January 2010
Families of zinc metalloproteases October 1994
Translocon Declogger Ste24 Protects against IAPP Oligomer-Induced Proteotoxicity March 2018
Phaser crystallographic software July 2007
Spectrometric Evaluation of the Approximate pK of the Carboxyl Group in 2,4-Dinitrophenyl-Amino Acids January 1962

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