Crystal Structure of a Pantothenate Synthetase from M. tuberculosis in complex with AMPCPP and pantoate

TB Structural Genomics Consortium (TBSGC)

DIPHOSPHOMETHYLPHOSPHONIC ACID ADENOSYL ESTER

Crystal structures of a pantothenate
synthetase from M. tuberculosis and its
complexes with substrates and a
reaction intermediate

Pantoate--beta-alanine ligase, Pantoate activating enzyme

PEG3000, lithium sulfate, magnesium sulfate, imidazole, ethanol, glycerol, pH 8.0, VAPOR DIFFUSION, HANGING DROP, temperature 293K

apo enzyme structure of M. tuberculosis pantothenate synthetase

CRYSTAL STRUCTURE OF A PANTOTHENATE SYNTHETASE FROM M. TUBERCULOSIS IN COMPLEX WITH AMPCPP AND PANTOATE (BETTER OCCUPANCY OF PANTOATE MOLECULE THAN 1N2E)

crystal structure of a pantothenate synthetase in complex with AMPCPP

CRYSTAL STRUCTURE OF A PANTOTHENATE SYNTHETASE FROM M. TUBERCULOSIS IN COMPLEX WITH A REACTION INTERMEDIATE, PANTOYL ADENYLATE

CRYSTAL STRUCTURE OF PANTOTHENATE SYNTHETASE FROM M. TUBERCULOSIS IN COMPLEX WITH A REACTION INTERMEDIATE, PANTOYL ADENYLATE, DIFFERENT OCCUPANCIES OF PANTOYL ADENYLATE.

crystal structure of a pantothenate synthetase from M. tuberculosis in complex with pantoate

CRYSTAL STRUCTURE OF PANTOTHENATE SYNTHETASE FROM M. TUBERCULOSIS, LOW OCCUPANCY OF BETA-ALANINE AT THE PANTOATE BINDING SITES

HETEROGEN
Entry 1N2B and this entry (1N2E) are two
different experiments that together indicate
that AMPCPP and pantoate cannot occupy the same
active site simultaneously. The crystal for
entry 1N2E was soaked in pantoate and AMPCPP
overnight, and partial occupancy of AMPCPP and
pantoate in subunit A was observed. The crystal
for entry 1N2B was soaked in twice concentration
of pantoate and same concentration of AMPCPP for
over 24 hours, and this gave higher occupancy of
pantoate, but lower occupancy of AMPCPP in
subunit A. Both entries have full occupancy of
AMPCPP in subunit B with a glycerol at the
pantoate binding site. The structural difference
between the two entries is higher occupancy of
pantoate and lower occupancy of AMPCPP in subunit
A of 1N2B relative to that of 1N2E.

SEQUENCE
RESIDUE 2 IS AN ALANINE IN THE SEQUENCE.
MUTATION OF THIS RESIDUE WAS NECESSARY TO
GENERATE AN NCOI RESTRICTION SITE
FOR CLONING INTO THE EXPRESSION VECTOR PET30A.

THE LAST 9 RESIDUES AFTER ARG300 WERE CLEAVED
OFF BY ENTEROKINASE DIGESTION (UNINTENTIONALLY),
WHICH WAS CARRIED OUT TO CLEAVE THE N-TERMINAL
TAG FROM THE RECOMBINANT PROTEIN.

full occupancy of AMPCPP in subunit B, partial occupancy of AMPCPP and pantoate in subunit A, at 1.6 A resolution.

Structural genomics, Rossmann fold, dimer, intersubunit beta sheet, PSI, Protein Structure Initiative, TB Structural Genomics Consortium, TBSGC, LIGASE