PDBsum entry 2cli

Lyase

PDB id: 2cli

Contents

Protein chains

258 a.a. *

395 a.a. *

Ligands

F9F

PLP

Metals

_NA

Waters ×591

* Residue conservation analysis

PDB id:

2cli

Name:

Lyase

Title:

Tryptophan synthase in complex with n-(4'- trifluoromethoxybenzenesulfonyl)-2-amino-1-ethylphosphate (f9)

Structure:

Tryptophan synthase alpha chain. Chain: a. Engineered: yes. Tryptophan synthase beta chain. Chain: b. Engineered: yes. Other_details: internal aldimine formed between b-k87 and c4 of pyridoxal-5'-phosphate

Source:

Salmonella typhimurium. Organism_taxid: 602. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

1.70Å R-factor: 0.188 R-free: 0.206

Authors:

H.Ngo,R.Harris,N.Kimmich,P.Casino,D.Niks,L.Blumenstein,T.R.Barends, V.Kulik,M.Weyand,I.Schlichting,M.F.Dunn

Key ref:

H.Ngo et al. (2007). Synthesis and characterization of allosteric probes of substrate channeling in the tryptophan synthase bienzyme complex. Biochemistry, 46, 7713-7727. PubMed id: 17559195

Date:

27-Apr-06 Release date: 12-Jun-07

Protein chain

P00929  (TRPA_SALTY) -  Tryptophan synthase alpha chain from Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)

Seq: 268 a.a.

Struc: 258 a.a.

Protein chain

P0A2K1  (TRPB_SALTY) -  Tryptophan synthase beta chain from Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)

Seq: 397 a.a.

Struc: 395 a.a.

Enzyme reactions

• Enzyme class: Chains A, B: E.C.4.2.1.20 - tryptophan synthase.
• Pathway: Tryptophan Biosynthesis
• Reaction: (1S,2R)-1-C-(indol-3-yl)glycerol 3-phosphate + L-serine = D-glyceraldehyde 3-phosphate + L-tryptophan + H2O
• Cofactor: Pyridoxal 5'-phosphate

Pyridoxal 5'-phosphate (Bound ligand (Het Group name = PLP) matches with 93.75% similarity)

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

Biochemistry 46:7713-7727 (2007)

PubMed id: 17559195

Synthesis and characterization of allosteric probes of substrate channeling in the tryptophan synthase bienzyme complex.

H.Ngo, R.Harris, N.Kimmich, P.Casino, D.Niks, L.Blumenstein, T.R.Barends, V.Kulik, M.Weyand, I.Schlichting, M.F.Dunn.

ABSTRACT

Allosteric interactions regulate substrate channeling in Salmonella typhimurium tryptophan synthase. The channeling of indole between the alpha- and beta-sites via the interconnecting 25 A tunnel is regulated by allosteric signaling arising from binding of ligand to the alpha-site, and covalent reaction of l-Ser at the beta-site. This signaling switches the alpha- and beta-subunits between open conformations of low activity and closed conformations of high activity. Our objective is to synthesize and characterize new classes of alpha-site ligands (ASLs) that mimic the binding of substrates, 3-indole-d-glycerol 3'-phosphate (IGP) or d-glyceraldehyde 3-phosphate (G3P), for use in the investigation of alpha-site-beta-site interactions. The new synthesized IGP analogues contain an aryl group linked to an O-phosphoethanolamine moiety through amide, sulfonamide, or thiourea groups. The G3P analogue, thiophosphoglycolohydroxamate, contains a hydroxamic acid group linked to a thiophosphate moiety. Crystal structures of the internal aldimine complexed with G3P and with three of the new ASLs are presented. These structural and solution studies of the ASL complexes with the internal aldimine form of the enzyme establish the following. (1) ASL binding occurs with high specificity and relatively high affinities at the alpha-site. (2) Binding of the new ASLs slows the entry of indole analogues into the beta-site by blocking the tunnel opening at the alpha-site. (3) ASL binding stabilizes the closed conformations of the beta-subunit for the alpha-aminoacrylate and quinonoid forms of the enzyme. (4) The new ASLs exhibit allosteric properties that parallel the behaviors of IGP and G3P.