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

0557998 - MBÚ 2023 RIV US eng J - Journal Article
Giladi, M. - Bar-El, M. - Vaňková, Pavla - Ferofontov, A. - Melvin, E. - Alkaderi, S. - Kavan, Daniel - Redko, B. - Haimov, E. - Wiener, R. - Man, Petr - Haitin, Y.
Structural basis for long-chain isoprenoid synthesis by cis-prenyltransferases.
Science Advances. Roč. 8, č. 20 (2022), č. článku eabn1171. ISSN 2375-2548. E-ISSN 2375-2548
R&D Projects: GA MŠMT(CZ) LM2018127; GA MŠMT(CZ) ED1.1.00/02.0109
EU Projects: European Commission(XE) 731077 - EU_FT-ICR_MS
Institutional support: RVO:61388971
Keywords : undecaprenyl-pyrophosphate synthase * length determination mechanism * nogo-b receptor * protein glycosylation * exchange * inhibitor * insights * mutation * subunit * complex
OECD category: Biochemistry and molecular biology
Impact factor: 13.6, year: 2022
Method of publishing: Open access
https://www.science.org/doi/10.1126/sciadv.abn1171

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.
Permanent Link: http://hdl.handle.net/11104/0331948