3KSL

Structure of FPT bound to DATFP-DH-GPP

Summary for 3KSL

• Entry DOI: 10.2210/pdb3ksl/pdb
• Descriptor: Farnesyltransferase, CAAX box, alpha, Protein farnesyltransferase subunit beta, MAGNESIUM ION, ... (6 entities in total)
• Functional Keywords: transferase, prenyltransferase, metal-binding, phosphoprotein, zinc
• Biological source: Rattus norvegicus (brown rat,rat,rats); More
• Total number of polymer chains: 2
• Total formula weight: 93370.37

Authors

Hovlid, M.L.,Edelstein, R.L.,Henry, O.,Ochocki, J.,DeGraw, A.,Lenevich, S.,Talbot, T.,Young, V.,Hruza, A.W.,Lopez-Gallego, F.,Labello, N.P.,Strickland, C.L.,Schmidt-Dannert, C.,Distefano, M.D. (deposition date: 2009-11-23, release date: 2009-12-22, Last modification date: 2023-09-06)

Primary citation

Hovlid, M.L.,Edelstein, R.L.,Henry, O.,Ochocki, J.,DeGraw, A.,Lenevich, S.,Talbot, T.,Young, V.G.,Hruza, A.W.,Lopez-Gallego, F.,Labello, N.P.,Strickland, C.L.,Schmidt-Dannert, C.,Distefano, M.D.
Synthesis, properties, and applications of diazotrifluropropanoyl-containing photoactive analogs of farnesyl diphosphate containing modified linkages for enhanced stability.
Chem.Biol.Drug Des., 75:51-67, 2010

PubMed Abstract: Photoactive analogs of farnesyl diphosphate (FPP) are useful probes in studies of enzymes that employ this molecule as a substrate. Here, we describe the preparation and properties of two new FPP analogs that contain diazotrifluoropropanoyl photophores linked to geranyl diphosphate via amide or ester linkages. The amide-linked analog (3) was synthesized in 32P-labeled form from geraniol in seven steps. Experiments with Saccharomyces cerevisiae protein farnesyltransferase (ScPFTase) showed that 3 is an alternative substrate for the enzyme. Photolysis experiments with [(32)P]3 demonstrate that this compound labels the beta-subunits of both farnesyltransferase and geranylgeranyltransferase (types 1 and 2). However, the amide-linked probe 3 undergoes a rearrangement to a photochemically unreactive isomeric triazolone upon long term storage making it inconvenient to use. To address this stability issue, the ester-linked analog 4 was prepared in six steps from geraniol. Computational analysis and X-ray crystallographic studies suggest that 4 binds to protein farnesyl transferase (PFTase) in a similar fashion as FPP. Compound 4 is also an alternative substrate for PFTase, and a 32P-labeled form selectively photocrosslinks the beta-subunit of ScPFTase as well as E. coli farnesyldiphosphate synthase and a germacrene-producing sesquiterpene synthase from Nostoc sp. strain PCC7120 (a cyanobacterial source). Finally, nearly exclusive labeling of ScPFTase in crude E. coli extract was observed, suggesting that [32P]4 manifests significant selectivity and should hence be useful for identifying novel FPP-utilizing enzymes in crude protein preparations.

PubMed: 19954434
DOI: 10.1111/j.1747-0285.2009.00914.x

Experimental method

X-RAY DIFFRACTION (2.05 Å)