Catalytic properties of the integral membrane RAS protease Rce1
- 1Structural Biology Program, Sloan Kettering Institute, Memorial Sloan Kettering Cancer Center, New York, New York, USA
- 2Louis V. Gerstner Jr. Graduate School of Biomedical Sciences, Memorial Sloan Kettering Cancer Center, New York, New York, USA
- 3Graduate Program in Biochemistry and Structural Biology, Cell and Developmental Biology, and Molecular Biology, Weill Cornell Medicine Graduate School of Medical Sciences, New York, New York, USA
Abstract
The integral membrane protease Rce1 cleaves prenylated CaaX proteins, including all RAS GTPases, as part of their posttranslational maturation. This cleavage, which removes the -aaX residues, is necessary for the proper cellular location and signaling properties of RAS and its oncogenic forms. Poor biochemical stabilities of yeast and mammalian Rce1 have hindered the purification of Rce1 in active form and limited studies addressing its catalytic properties. Here, we identify that Drosophila melanogaster Rce1 has good biochemical stability and is catalytically active when purified. The purified protein recapitulates known properties of Rce1, including specificity for prenylated CaaX substrates. Like other integral membrane proteases, the enzyme has a low turnover number (kcat), of approximately 20 h−1. Pre-steady-state kinetic analysis reveals that product release is the rate-limiting step. Our data indicate that Rce1 is inhibited by nanomolar levels of Zn2+, but that metals are not involved in catalysis. The data suggest that Zn2+ is a competitive inhibitor with respect to the substrate. The signaling lipid PA also inhibits the enzyme in vitro, raising the possibility that the cellular activity of Rce1 may be modulated by lipids and/or Zn2+. These studies combined with structural predictions and prior investigation lead to the hypothesis that the active site contains a catalytic Glu-His-His triad of amino acids that coordinate the substrate and form the inhibitory Zn2+ site. This work enhances the understanding of the enzyme and gives insight into means for Rce1 inhibition.
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BibTeX
@article{dongen2026catalytic,
title = {{Catalytic properties of the integral membrane RAS protease Rce1}},
author = {David J. Van Dongen and Abbie M. Collette and Melinda M. Diver and Stephen B. Long},
journal = {The Journal of Biological Chemistry},
year = {2026},
month = sep,
doi = {10.1016/j.jbc.2026.113525},
url = {https://doi.org/10.1016/j.jbc.2026.113525},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Catalytic properties of the integral membrane RAS protease Rce1
AU - David J. Van Dongen
AU - Abbie M. Collette
AU - Melinda M. Diver
AU - Stephen B. Long
PY - 2026
DA - 2026/09/08
JO - The Journal of Biological Chemistry
DO - 10.1016/j.jbc.2026.113525
UR - https://doi.org/10.1016/j.jbc.2026.113525
AB - The integral membrane protease Rce1 cleaves prenylated CaaX proteins, including all RAS GTPases, as part of their posttranslational maturation. This cleavage, which removes the -aaX residues, is necessary for the proper cellular location and signaling properties of RAS and its oncogenic forms. Poor biochemical stabilities of yeast and mammalian Rce1 have hindered the purification of Rce1 in active form and limited studies addressing its catalytic properties. Here, we identify that Drosophila melanogaster Rce1 has good biochemical stability and is catalytically active when purified. The purified protein recapitulates known properties of Rce1, including specificity for prenylated CaaX substrates. Like other integral membrane proteases, the enzyme has a low turnover number (kcat), of approximately 20 h−1. Pre-steady-state kinetic analysis reveals that product release is the rate-limiting step. Our data indicate that Rce1 is inhibited by nanomolar levels of Zn2+, but that metals are not involved in catalysis. The data suggest that Zn2+ is a competitive inhibitor with respect to the substrate. The signaling lipid PA also inhibits the enzyme in vitro, raising the possibility that the cellular activity of Rce1 may be modulated by lipids and/or Zn2+. These studies combined with structural predictions and prior investigation lead to the hypothesis that the active site contains a catalytic Glu-His-His triad of amino acids that coordinate the substrate and form the inhibitory Zn2+ site. This work enhances the understanding of the enzyme and gives insight into means for Rce1 inhibition.
SN - 0021-9258
SN - 1083-351X
ER -