Sequence-specific 1H, 13C and 15N resonance assignments of rat liver
fructose-2,6-bisphosphatase domain

The molecule studied here contains 30 additional residues at the C-terminus.

The molecule studied here contains 30 additional residues at the C-terminus,
and it is not a phospho-histidine intermediate.

The system studied here is the biphosphatase of the bifunctional enzyme only.
and it is from rat liver instead of rat testis.

For residues 219-221 a second set of resonances with slightly different
chemical shifts was observed.

Sequence-specific 1H, 13C and 15N resonance assignments of the rat liver fructose-2,6-bisphosphatase domain.

The NMRPipe system is a UNIX software environment of processing, graphics, and
analysis tools designed to meet current routine and research-oriented
multidimensional processing requirements, and to anticipate and accommodate
future demands and developments. The system is based on UNIX pipes, which allow
programs running simultaneously to exchange streams of data under user control.
In an NMRPipe processing scheme, a stream of spectral data flows through a
pipeline of processing programs, each of which performs one component of the
overall scheme, such as Fourier transformation or linear prediction. Complete
multidimensional processing schemes are constructed as simple UNIX shell
scripts. The processing modules themselves maintain and exploit accurate
records of data sizes, detection modes, and calibration information in all
dimensions, so that schemes can be constructed without the need to explicitly
define or anticipate data sizes or storage details of real and imaginary
channels during processing. The asynchronous pipeline scheme provides other
substantial advantages, including high flexibility, favorable processing
speeds, choice of both all-in-memory and disk-bound processing, easy adaptation
to different data formats, simpler software development and maintenance, and
the ability to distribute processing tasks on multi-CPU computers and computer
networks.

Delaglio F, Grzesiek S, Vuister GW, Zhu G, Pfeifer J, Bax A.
NMRPipe: a multidimensional spectral processing system based on UNIX pipes.
J Biomol NMR. 1995 Nov;6(3):277-93.

NMRPipe: a multidimensional spectral processing system based on UNIX pipes.

Johnson, B. A., and Blevins, R. A. (1994)
J.Biomol.NMR 4, 603-614.

The crystal structure of the recombinant fructose-2,6-bisphosphatase domain,
which covers the residues between 251 and 440 of the rat liver bifunctional
enzyme, 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase, was determined by
multiwavelength anomalous dispersion phasing and refined at 2.5 A resolution.
The selenomethionine-substituted protein was induced in the methionine
auxotroph, Escherichia coli DL41DE3, purified, and crystallized in a manner
similar to that of the native protein. Phase information was calculated using
the multiwavelength anomalous dispersion data collected at the X-ray
wavelengths near the absorption edge of the K-shell alpha electrons of
selenium. The fructose-2,6-bisphosphatase domain has a core alpha/beta
structure which consists of six stacked beta-strands, four parallel and two
antiparallel. The core beta-sheet is surrounded by nine alpha-helices. The
catalytic site, as defined by a bound phosphate ion, is positioned near the
C-terminal end of the beta-sheet and close to the N-terminal end of an
alpha-helix. The active site pocket is funnel-shaped. The narrow opening of the
funnel is wide enough for a water molecule to pass. The key catalytic residues,
including His7, His141, and Glu76, are near each other at the active site and
probably function as general acids and/or bases during a catalytic cycle. The
inorganic phosphate molecule is bound to an anion trap formed by Arg6, His7,
Arg56, and His141. The core structure of the Fru-2,6-P2ase is similar to that
of the yeast phosphoglycerate mutase and the rat prostatic acid phosphatase.
However, the structure of one of the loops near the active site is completely
different from the other family members, perhaps reflecting functional
differences and the nanomolar range affinity of Fru-2,6-P2ase for its
substrate. The imidazole rings of the two key catalytic residues, His7 and
His141, are not parallel as in the yeast phosphoglycerate mutase. The crystal
structure is used to interpret the existing chemical data already available for
the bisphosphatase domain. In addition, the crystal structure is compared with
two other proteins that belong to the histidine phosphatase family.

Lee, Y. H., Ogata, C., Pflugrath, J. W., Levitt, D. G., Sarma, R.,
Banaszak, L. J., and Pilkis, S. J. (1996)
Biochemistry 35(19), 6010-9.

Crystal structure of the rat liver fructose-2,6-bisphosphatase based on selenomethionine multiwavelength anomalous dispersion phases.

BACKGROUND. Glucose homeostasis is maintained by the processes of glycolysis and
gluconeogenesis. The importance of these pathways is demonstrated by the severe
and life threatening effects observed in various forms of diabetes. The
bifunctional enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase
catalyzes both the synthesis and degradation of fructose-2,6-bisphosphate, a
potent regulator of glycolysis. Thus this bifunctional enzyme plays an indirect
yet key role in the regulation of glucose metabolism. RESULTS. We have
determined the 2.0 A crystal structure of the rat testis isozyme of this
bifunctional enzyme. The enzyme is a homodimer of 55 kDa subunits arranged in a
head-to-head fashion, with each monomer consisting of independent kinase and
phosphatase domains. The location of ATPgammaS and inorganic phosphate in the
kinase and phosphatase domains, respectively, allow us to locate and describe
the active sites of both domains. CONCLUSIONS. The kinase domain is clearly
related to the superfamily of mononucleotide binding proteins, with a
particularly close relationship to the adenylate kinases and the
nucleotide-binding portion of the G proteins. This is in disagreement with the
broad speculation that this domain would resemble phosphofructokinase. The
phosphatase domain is structurally related to a family of proteins which
includes the cofactor independent phosphoglycerate mutases and acid
phosphatases.

Hasemann, C. A., Istvan, E. S., Uyeda, K., and Deisenhofer, J. (1996)
Structure 4(9), 1017-29.

The crystal structure of the bifunctional enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase reveals distinct domain homologies.

Regulatory Complex Of Fructose-2,6-bisphosphatase

Michaelis Complex Of Fructose-2,6-bisphosphatase

The Bisphosphatase Domain Of The Bifunctional Rat Liver 6- Phosphofructo-2-KinaseFRUCTOSE-2,6-Bisphosphatase

The Bisphosphatase Domain Of The Bifunctional Rat Liver 6- Phosphofructo-2-kinase/fructose-2,6-bisphosphatase

unnamed protein product [Rattus norvegicus]

6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase [Rattus norvegicus]

6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase [Bos taurus]

6-phosphofructo 2-kinase/fructose 2, 6-bisphosphatase, partial [Rattus norvegicus]

fructose-2,6-bisphosphatase [Bos taurus]

6-phosphofructo-2-kinase (EC 2.7.1.105) / fructose-2, 6-bisphosphate 2-phosphatase (EC 3.1.3.46) clone 5c, skeletal muscle - ra

6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 1 isoform 2 [Rattus norvegicus]

6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 1 isoform 3 [Rattus norvegicus]

6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 1 isoform 2 [Homo sapiens]

6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 1 isoform 3 [Homo sapiens]

6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 1 isoform 1 [Homo sapiens]

RecName: Full=6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 1; Short=6PF-2-K/Fru-2,6-P2ase 1; Short=PFK/FBPase 1; AltNam

TPA: 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 1 [Bos taurus]

TROSY versions used for all experiments;

2H decoupling used on triply-labeled sample.

A cryo-probe was used on the Bruker Avance 600.

F.Delaglio, S.Grzesiek, G.W.Vuister, G.Zhu, J.Pfeifer and A.Bax,
J.Biomol. NMR 6, 277-293 (1995).

B.A.Johnson and R.A.Blevins
J.Biomol. NMR 4, 603-614 (1994).