7PAX

Structure of the human heterotetrameric cis-prenyltransferase complex in complex with magnesium, FsPP and IPP

Summary for 7PAX

• Entry DOI: 10.2210/pdb7pax/pdb
• Descriptor: Dehydrodolichyl diphosphate synthase complex subunit DHDDS, Dehydrodolichyl diphosphate synthase complex subunit NUS1, 3-METHYLBUT-3-ENYL TRIHYDROGEN DIPHOSPHATE, ... (6 entities in total)
• Functional Keywords: dhdds, ngbr, hcit, cis-prenyltransferase, dolichol, transferase
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 2
• Total formula weight: 64082.35

Authors

Giladi, M.,Lisnyansky Bar-El, M.,Haitin, Y. (deposition date: 2021-07-30, release date: 2022-06-01, Last modification date: 2024-01-31)

Primary citation

Giladi, M.,Lisnyansky Bar-El, M.,Vankova, P.,Ferofontov, A.,Melvin, E.,Alkaderi, S.,Kavan, D.,Redko, B.,Haimov, E.,Wiener, R.,Man, P.,Haitin, Y.
Structural basis for long-chain isoprenoid synthesis by cis -prenyltransferases.
Sci Adv, 8:eabn1171-eabn1171, 2022

PubMed 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 -prenyltransferase complex (h-PT), residing at the endoplasmic reticulum membrane and playing a crucial role in protein glycosylation. Crystallographic investigation of h-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.

PubMed: 35584224
DOI: 10.1126/sciadv.abn1171

Experimental method

X-RAY DIFFRACTION (2 Å)