Structural basis for long-chain isoprenoid synthesis by cis-prenyltransferases

Moshe Giladi*, Michal Lisnyansky Bar-El, Pavla Vaňková, Alisa Ferofontov, Emelia Melvin, Suha Alkaderi, Daniel Kavan, Boris Redko, Elvira Haimov, Reuven Wiener, Petr Man, Yoni Haitin

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Isoprenoids are synthesized by the prenyltransferase superfamily, which is subdivided according to the product stereoisomerism and length. In short- and medium-chain isoprenoids, product length correlates with active site volume. However, enzymes synthesizing long-chain products and rubber synthases fail to conform to this paradigm, because of an unexpectedly small active site. Here, we focused on the human cis-prenyltransferase complex (hcis-PT), residing at the endoplasmic reticulum membrane and playing a crucial role in protein glycosylation. Crystallographic investigation of hcis-PT along the reaction cycle revealed an outlet for the elongating product. Hydrogen-deuterium exchange mass spectrometry analysis showed that the hydrophobic active site core is flanked by dynamic regions consistent with separate inlet and outlet orifices. Last, using a fluorescence substrate analog, we show that product elongation and membrane association are closely correlated. Together, our results support direct membrane insertion of the elongating isoprenoid during catalysis, uncoupling active site volume from product length.

Original languageEnglish
Article numbereabn1171
JournalScience advances
Volume8
Issue number20
DOIs
StatePublished - May 2022

Funding

FundersFunder number
CIISBLM2018127
Claire and Amedee Maratier Institute for the Study of Blindness
Sackler Faculty of Medicine
Tel Aviv Medical Center, Israel
Visual Disorders
Israel Cancer Research Fund19202, 01214
European Synchrotron Radiation FacilityDHDDS.GW1, KG405
German-Israeli Foundation for Scientific Research and Development1775/12, I-2425-418.13/2016
Ministerstvo Školství, Mládeže a TělovýchovyCZ.1.05/1.1.00/02.0109
Israel Cancer Association20200037
Israel Science Foundation1653/21, 1721/16
Tel Aviv University
Horizon 2020731077

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