Mass spectrometry captures off-target drug binding and provides mechanistic insights into the human metalloprotease ZMPSTE24

Shahid Mehmood, Julien Marcoux, Joseph Gault, Andrew Quigley, Susan Doris Michaelis, Stephen G. Young, Elisabeth P. Carpenter, Carol V. Robinson

Research output: Contribution to journalArticle

Abstract

Off-target binding of hydrophobic drugs can lead to unwanted side effects, either through specific or non-specific binding to unintended membrane protein targets. However, distinguishing the binding of drugs to membrane proteins from that of detergents, lipids and cofactors is challenging. Here, we use high-resolution mass spectrometry to study the effects of HIV protease inhibitors on the human zinc metalloprotease ZMPSTE24. This intramembrane protease plays a major role in converting prelamin A to mature lamin A. We monitored the proteolysis of farnesylated prelamin A peptide by ZMPSTE24 and unexpectedly found retention of the C-terminal peptide product with the enzyme. We also resolved binding of zinc, lipids and HIV protease inhibitors and showed that drug binding blocked prelamin A peptide cleavage and conferred stability to ZMPSTE24. Our results not only have relevance for the progeria-like side effects of certain HIV protease inhibitor drugs, but also highlight new approaches for documenting off-target drug binding.

Original languageEnglish (US)
Pages (from-to)1152-1158
Number of pages7
JournalNature Chemistry
Volume8
Issue number12
DOIs
StatePublished - Dec 1 2016

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Metalloproteases
Mass spectrometry
HIV Protease Inhibitors
Peptides
Pharmaceutical Preparations
Lipids
Zinc
Proteolysis
Proteins
Membranes
Membrane Proteins
Lamin Type A
Detergents
Enzymes
Peptide Hydrolases
prelamin A

ASJC Scopus subject areas

  • Chemistry(all)
  • Chemical Engineering(all)

Cite this

Mass spectrometry captures off-target drug binding and provides mechanistic insights into the human metalloprotease ZMPSTE24. / Mehmood, Shahid; Marcoux, Julien; Gault, Joseph; Quigley, Andrew; Michaelis, Susan Doris; Young, Stephen G.; Carpenter, Elisabeth P.; Robinson, Carol V.

In: Nature Chemistry, Vol. 8, No. 12, 01.12.2016, p. 1152-1158.

Research output: Contribution to journalArticle

Mehmood, S, Marcoux, J, Gault, J, Quigley, A, Michaelis, SD, Young, SG, Carpenter, EP & Robinson, CV 2016, 'Mass spectrometry captures off-target drug binding and provides mechanistic insights into the human metalloprotease ZMPSTE24', Nature Chemistry, vol. 8, no. 12, pp. 1152-1158. https://doi.org/10.1038/nchem.2591
Mehmood, Shahid ; Marcoux, Julien ; Gault, Joseph ; Quigley, Andrew ; Michaelis, Susan Doris ; Young, Stephen G. ; Carpenter, Elisabeth P. ; Robinson, Carol V. / Mass spectrometry captures off-target drug binding and provides mechanistic insights into the human metalloprotease ZMPSTE24. In: Nature Chemistry. 2016 ; Vol. 8, No. 12. pp. 1152-1158.
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