PDBsum entry 4x1k

Structural protein/inhibitor

PDB id: 4x1k

Contents

Protein chains

429 a.a.

428 a.a.

123 a.a.

Ligands

GTP ×2

GDP ×2

LOC ×2

3WZ ×2

Metals

_MG ×2

PDB id:

4x1k

Name:

Structural protein/inhibitor

Title:

Discovery of cytotoxic dolastatin 10 analogs with n-terminal modifications

Structure:

Tubulin alpha chain. Chain: a, c. Tubulin beta chain. Chain: b, d. Stathmin-4. Chain: e. Fragment: unp residues 49-189. Synonym: stathmin-like protein b3,rb3. Engineered: yes

Source:

Ovis aries. Sheep. Organism_taxid: 9940. Rattus norvegicus. Rat. Organism_taxid: 10116. Gene: stmn4. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

3.50Å R-factor: 0.203 R-free: 0.263

Authors:

K.D.Parris

Key ref:

A.Maderna et al. (2014). Discovery of cytotoxic dolastatin 10 analogues with N-terminal modifications. J Med Chem, 57, 10527-10543. PubMed id: 25431858 DOI: 10.1021/jm501649k

Date:

24-Nov-14 Release date: 25-Mar-15

Protein chains

D0VWZ0  (D0VWZ0_SHEEP) -  Tubulin alpha chain from Ovis aries

Seq: 451 a.a.

Struc: 429 a.a.

Protein chains

D0VWY9  (D0VWY9_SHEEP) -  Tubulin beta chain from Ovis aries

Seq: 445 a.a.

Struc: 428 a.a.

Protein chain

P63043  (STMN4_RAT) -  Stathmin-4 from Rattus norvegicus

Seq: 189 a.a.

Struc: 123 a.a.*

* PDB and UniProt seqs differ at 2 residue positions (black crosses)

DOI no: 10.1021/jm501649k

J Med Chem 57:10527-10543 (2014)

PubMed id: 25431858

Discovery of cytotoxic dolastatin 10 analogues with N-terminal modifications.

A.Maderna, M.Doroski, C.Subramanyam, A.Porte, C.A.Leverett, B.C.Vetelino, Z.Chen, H.Risley, K.Parris, J.Pandit, A.H.Varghese, S.Shanker, C.Song, S.C.Sukuru, K.A.Farley, M.M.Wagenaar, M.J.Shapiro, S.Musto, M.H.Lam, F.Loganzo, C.J.O'Donnell.

ABSTRACT

Auristatins, synthetic analogues of the antineoplastic natural product Dolastatin 10, are ultrapotent cytotoxic microtubule inhibitors that are clinically used as payloads in antibody-drug conjugates (ADCs). The design and synthesis of several new auristatin analogues with N-terminal modifications that include amino acids with α,α-disubstituted carbon atoms are described, including the discovery of our lead auristatin, PF-06380101. This modification of the peptide structure is unprecedented and led to analogues with excellent potencies in tumor cell proliferation assays and differential ADME properties when compared to other synthetic auristatin analogues that are used in the preparation of ADCs. In addition, auristatin cocrystal structures with tubulin are being presented that allow for the detailed examination of their binding modes. A surprising finding is that all analyzed analogues have a cis-configuration at the Val-Dil amide bond in their functionally relevant tubulin bound state, whereas in solution this bond is exclusively in the trans-configuration. This remarkable observation shines light onto the preferred binding mode of auristatins and serves as a valuable tool for structure-based drug design.