Geranylgeranyl transferase type I (GGTase-I) structure and uses thereof
Protein geranylgeranyl transferase type I (GGTase-I), a Ca 1 a 2 X prenyltransferase, is an essential enzyme in eukaryotes. GGTase-I catalyzes the carboxyl-terminal lipidation of over one hundred proteins, including many GTP-binding regulatory proteins (G proteins). The presently disclosed subject matter comprises a plurality of crystal structures of mammalian GGTase-I in complex with substrates and products that provide the first structural information for this enzyme, facilitating the elucidation of a generalized method of action for all protein prenyltransferases; the method includes a role in product transport. The structures reveal specificity determinants that allow classification of putative protein prenyltransferase sequences and can facilitate optimization of GGTase-I and FTase modulators.
1. A method for designing a ligand for a farnesyl transferase (ETase), the method comprising:
(a) providing a molecule that binds to a geranylgeranyl transferase type I (GGTase-l) a 2 site;
(b) determining a moiety of the molecule that interacts with the GGTase-I a 2 site;
(c) modifying the moiety to produce a modified molecule; wherein the modifying comprises modifying a moiety that interacts with one or more of residues Thr491β, Phe 106β, and Leu 361β of a human GGTase-I polypeptide;
(d) modeling a first interaction between the modified molecule and the GGTase-I a 2 sites
(e) modeling a second interaction between the modified molecule with an FTase a 2 site;
(f) identifying in a modified molecule that is predicted to be unable to bind to the GGTase-I a 2 site but able to bind to the FTase a 2 site; and
(g) displaying a structure of the modified molecule identified in step (f),
whereby a ligand for a FTase is designed.
2. The method of claim 1 , wherein the ligand comprises an inhibitor.
3. The method of claim 1 , wherein the determining comprises identifying a moiety of the molecule that interacts with residues Thr 49β, Phe 106β, and Leu 361β of a human GGTase-I polypeptide.
4. The method of claim 1 , wherein the modifying comprises substituting the moiety with a group comprising an aromatic ring.
5. The method of claim 1 , wherein the molecule is a peptide or a small molecule.
6. The method of claim 5 , wherein the modified molecule comprises a change in an amino acid that interacts with the a 2 site of the GGTase from an amino acid with a non-aromatic side chain to an amino acid comprising an aromatic side chain.
7. The method of claim 1 , wherein the modeling a first interaction between the modified molecule and the GGTase-l a 2 site comprises evaluating the first interaction between the modified molecule and the GGTase-l a 2 site with respect to a three-dimensional structure of a crystal comprising the GGTase-l a 2 site. aromatic side chain.