1WCB

PLP-DEPENDENT CATALYTIC ANTIBODY 15A9 IN COMPLEX WITH ITS HAPTEN

Summary for 1WCB

• Entry DOI: 10.2210/pdb1wcb/pdb
• Related: 1WC7 2BMK
• Descriptor: FAB FRAGMENT OF CATALYTIC ANTIBODY 15A9, LIGHT CHAIN, FAB FRAGMENT OF CATALYTIC ANTIBODY 15A9, HEAVY CHAIN, N~2~-({3-HYDROXY-2-METHYL-5-[(PHOSPHONOOXY)METHYL]PYRIDIN-4-YL}METHYL)-L-LYSINE, ... (5 entities in total)
• Functional Keywords: catalytic antibody, transamination, pyridoxal-phosphate, hapten, phosphopyridoxyl-l-lysine, immune system
• Biological source: MUS MUSCULUS (MOUSE); More
• Total number of polymer chains: 4
• Total formula weight: 97611.27

Authors

Golinelli-Pimpaneau, B.,Christen, P. (deposition date: 2004-11-12, release date: 2006-03-03, Last modification date: 2024-10-23)

Primary citation

Golinelli-Pimpaneau, B.,Luthi, C.,Christen, P.
Structural Basis for D-Amino Acid Transamination by the Pyridoxal- 5' -Phosphate - Dependent Catalytic Antibody 15A9.
J.Biol.Chem., 281:23969-, 2006

PubMed Abstract: Antibody 15A9, raised with 5'-phosphopyridoxyl (PPL)-N(epsilon)-acetyl-L-lysine as hapten, catalyzes the reversible transamination of hydrophobic D-amino acids with pyridoxal 5'-phosphate (PLP). The crystal structures of the complexes of Fab 15A9 with PPL-L-alanine, PPL-D-alanine, and the hapten were determined at 2.3, 2.3, and 2.5A resolution, respectively, and served for modeling the complexes with the corresponding planar imine adducts. The conformation of the PLP-amino acid adduct and its interactions with 15A9 are similar to those occurring in PLP-dependent enzymes, except that the amino acid substrate is only weakly bound, and, due to the immunization and selection strategy, the lysine residue that covalently binds PLP in these enzymes is missing. However, the N-acetyl-L-lysine moiety of the hapten appears to have selected for aromatic residues in hypervariable loop H3 (Trp-H100e and Tyr-H100b), which, together with Lys-H96, create an anion-binding environment in the active site. The structural situation and mutagenesis experiments indicate that two catalytic residues facilitate the transamination reaction of the PLP-D-alanine aldimine. The space vacated by the absent L-lysine side chain of the hapten can be filled, in both PLP-alanine aldimine complexes, by mobile Tyr-H100b. This group can stabilize a hydroxide ion, which, however, abstracts the C alpha proton only from D-alanine. Together with the absence of any residue capable of deprotonating C alpha of L-alanine, Tyr-H100b thus underlies the enantiomeric selectivity of 15A9. The reprotonation of C4' of PLP, the rate-limiting step of 15A9-catalyzed transamination, is most likely performed by a water molecule that, assisted by Lys-H96, produces a hydroxide ion stabilized by the anion-binding environment.

PubMed: 16790434
DOI: 10.1074/JBC.M602184200

Experimental method

X-RAY DIFFRACTION (2.5 Å)