Structure, function and inhibition of the two- and three-domain 4Fe-4S IspG proteins

Yi Liang Liu, Francisco Guerra, Ke Wang, Weixue Wang, Jikun Li, Cancan Huang, Wei Zhu, Kevin Houlihan, Zhi Li, Yong Zhang, Satish K. Nair, Eric Oldfield

Research output: Contribution to journalArticlepeer-review

Abstract

IspG is a 4Fe4S protein involved in isoprenoid biosynthesis. Most bacterial IspGs contain two domains: a TIM barrel (A) and a 4Fe4S domain (B), but in plants and malaria parasites, there is a large insert domain (A*) whose structure and function are unknown. We show that bacterial IspGs function in solution as (AB)2 dimers and that mutations in either both A or both B domains block activity. Chimeras harboring an A-mutation in one chain and a B-mutation in the other have 50% of the activity seen in wild-type protein, because there is still one catalytically active AB domain. However, a plant IspG functions as an AA*B monomer. We propose, using computational modeling and electron microscopy, that the A*insert domain has a TIM barrel structure that interacts with the A domain. This structural arrangement enables the A and B domains to interact in a "cup and ball" manner during catalysis, just as in the bacterial systems. EPR/HYSCORE spectra of reaction intermediate, product, and inhibitor ligands bound to both two and three domain proteins are identical, indicating the same local electronic structure, and computational docking indicates these ligands bridge both A and B domains. Overall, the results are of broad general interest because they indicate the insert domain in three-domain IspGs is a second TIM barrel that plays a structural role and that the pattern of inhibition of both two and three domain proteins are the same, results that can be expected to be of use in drug design.

Original languageEnglish (US)
Pages (from-to)8558-8563
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume109
Issue number22
DOIs
StatePublished - May 29 2012

Keywords

  • Drug discovery
  • Iron-sulfur
  • Metalloprotein
  • Protein folding

ASJC Scopus subject areas

  • General

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