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References listed in PDB file
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Key reference
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Title
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Arginine 54 in the active site of escherichia coli aspartate transcarbamoylase is critical for catalysis: a site-Specific mutagenesis, Nmr, And X-Ray crystallographic study.
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Authors
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J.W.Stebbins,
D.E.Robertson,
M.F.Roberts,
R.C.Stevens,
W.N.Lipscomb,
E.R.Kantrowitz.
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Ref.
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Protein Sci, 1992,
1,
1435-1446.
[DOI no: ]
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PubMed id
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Abstract
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The replacement of Arg-54 by Ala in the active site of Escherichia coli
aspartate transcarbamoylase causes a 17,000-fold loss of activity but does not
significantly influence the binding of substrates or substrate analogs
(Stebbins, J.W., Xu, W., & Kantrowitz, E.R., 1989, Biochemistry 28,
2592-2600). In the X-ray structure of the wild-type enzyme, Arg-54 interacts
with both the anhydride oxygen and a phosphate oxygen of carbamoyl phosphate
(CP) (Gouaux, J.E. & Lipscomb, W.N., 1988, Proc. Natl. Acad. Sci. USA 85,
4205-4208). The Arg-54-->Ala enzyme was crystallized in the presence of the
transition state analog N-phosphonacetyl-L-aspartate (PALA), data were collected
to a resolution limit of 2.8 A, and the structure was solved by molecular
replacement. The analysis of the refined structure (R factor = 0.18) indicates
that the substitution did not cause any significant alterations to the active
site, except that the side chain of the arginine was replaced by two water
molecules. 31P-NMR studies indicate that the binding of CP to the wild-type
catalytic subunit produces an upfield chemical shift that cannot reflect a
significant change in the ionization state of the CP but rather indicates that
there are perturbations in the electronic environment around the phosphate
moiety when CP binds to the enzyme. The pH dependence of this upfield shift for
bound CP indicates that the catalytic subunit undergoes a conformational change
with a pKa approximately 7.7 upon CP binding. Furthermore, the linewidth of the
31P signal of CP bound to the Arg-54-->Ala enzyme is significantly narrower
than that of CP bound to the wild-type catalytic subunit at any pH, although the
change in chemical shift for the CP bound to the mutant enzyme is unaltered.
31P-NMR studies of PALA complexed to the wild-type catalytic subunit indicate
that the phosphonate group of the bound PALA exists as the dianion at pH 7.0 and
8.8, whereas in the Arg-54-->Ala catalytic subunit the phosphonate group of
the bound PALA exists as the monoanion at pH 7.0 and 8.8. Thus, the side chain
of Arg-54 is essential for the proper ionization of the phosphonate group of
PALA and by analogy the phosphate group in the transition state. These data
support the previously proposed proton transfer mechanism, in which a fully
ionized phosphate group in the transition state accepts a proton during
catalysis.
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Secondary reference #1
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Title
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Structural consequences of effector binding to the t state of aspartate carbamoyltransferase: crystal structures of the unligated and ATP- And ctp-Complexed enzymes at 2.6-A resolution.
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Authors
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R.C.Stevens,
J.E.Gouaux,
W.N.Lipscomb.
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Ref.
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Biochemistry, 1990,
29,
7691-7701.
[DOI no: ]
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PubMed id
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Secondary reference #2
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Title
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Crystal structures of aspartate carbamoyltransferase ligated with phosphonoacetamide, Malonate, And ctp or ATP at 2.8-A resolution and neutral ph.
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Authors
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J.E.Gouaux,
R.C.Stevens,
W.N.Lipscomb.
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Ref.
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Biochemistry, 1990,
29,
7702-7715.
[DOI no: ]
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PubMed id
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Secondary reference #3
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Title
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Crystal structures of phosphonoacetamide ligated t and phosphonoacetamide and malonate ligated r states of aspartate carbamoyltransferase at 2.8-A resolution and neutral ph.
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Authors
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J.E.Gouaux,
W.N.Lipscomb.
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Ref.
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Biochemistry, 1990,
29,
389-402.
[DOI no: ]
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PubMed id
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Secondary reference #4
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Title
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Structural transitions in crystals of native aspartate carbamoyltransferase.
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Authors
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J.E.Gouaux,
W.N.Lipscomb.
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Ref.
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Proc Natl Acad Sci U S A, 1989,
86,
845-848.
[DOI no: ]
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PubMed id
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Secondary reference #5
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Title
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Structure of a single amino acid mutant of aspartate carbamoyltransferase at 2.5-A resolution: implications for the cooperative mechanism.
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Authors
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J.E.Gouaux,
W.N.Lipscomb,
S.A.Middleton,
E.R.Kantrowitz.
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Ref.
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Biochemistry, 1989,
28,
1798-1803.
[DOI no: ]
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PubMed id
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Secondary reference #6
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Title
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Escherichia coli aspartate transcarbamylase: the relation between structure and function.
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Authors
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E.R.Kantrowitz,
W.N.Lipscomb.
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Ref.
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Science, 1988,
241,
669-674.
[DOI no: ]
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PubMed id
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Secondary reference #7
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Title
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Three-Dimensional structure of carbamoyl phosphate and succinate bound to aspartate carbamoyltransferase.
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Authors
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J.E.Gouaux,
W.N.Lipscomb.
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Ref.
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Proc Natl Acad Sci U S A, 1988,
85,
4205-4208.
[DOI no: ]
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PubMed id
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Secondary reference #8
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Title
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Structural asymmetry in the ctp-Liganded form of aspartate carbamoyltransferase from escherichia coli.
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Authors
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K.H.Kim,
Z.X.Pan,
R.B.Honzatko,
H.M.Ke,
W.N.Lipscomb.
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Ref.
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J Mol Biol, 1987,
196,
853-875.
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PubMed id
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Secondary reference #9
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Title
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2.5 a structure of aspartate carbamoyltransferase complexed with the bisubstrate analog n-(Phosphonacetyl)-L-Aspartate.
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Authors
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K.L.Krause,
K.W.Volz,
W.N.Lipscomb.
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Ref.
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J Mol Biol, 1987,
193,
527-553.
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PubMed id
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Secondary reference #10
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Title
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The catalytic mechanism of escherichia coli aspartate carbamoyltransferase: a molecular modelling study.
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Authors
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J.E.Gouaux,
K.L.Krause,
W.N.Lipscomb.
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Ref.
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Biochem Biophys Res Commun, 1987,
142,
893-897.
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PubMed id
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Secondary reference #11
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Title
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Structure at 2.9-A resolution of aspartate carbamoyltransferase complexed with the bisubstrate analogue n-(Phosphonacetyl)-L-Aspartate.
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Authors
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K.L.Krause,
K.W.Volz,
W.N.Lipscomb.
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Ref.
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Proc Natl Acad Sci U S A, 1985,
82,
1643-1647.
[DOI no: ]
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PubMed id
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Secondary reference #12
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Title
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Structure of unligated aspartate carbamoyltransferase of escherichia coli at 2.6-A resolution.
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Authors
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H.M.Ke,
R.B.Honzatko,
W.N.Lipscomb.
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Ref.
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Proc Natl Acad Sci U S A, 1984,
81,
4037-4040.
[DOI no: ]
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PubMed id
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Secondary reference #13
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Title
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Crystal and molecular structures of native and ctp-Liganded aspartate carbamoyltransferase from escherichia coli.
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Authors
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R.B.Honzatko,
J.L.Crawford,
H.L.Monaco,
J.E.Ladner,
B.F.Ewards,
D.R.Evans,
S.G.Warren,
D.C.Wiley,
R.C.Ladner,
W.N.Lipscomb.
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Ref.
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J Mol Biol, 1982,
160,
219-263.
[DOI no: ]
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PubMed id
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Figure 3.
FIG:. . Variation ofthtl crystallographic -factor with wsolution fi)r the 1132 (0) and 1'311 (m) rystal
forms. )ata ww grouped ccording o resolution in 20 hells venly spaced in limits of sine theta. The
solid inw rcpresrnt he t~hrorrtical variaton of th factor at he, sprcified co-ordiate error r.m.s.).
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Figure 10.
FIN: 10. Intrrfam betw-een catalytic chair) Cl (unbroken inrs) and rrgulatory and catalytic: chains H4
and 4 broken lines). Cl cont.rihutes Clu238 o a polar link ith Tyrl65 of C4. and thr tmrklwnr of
Ala237 and ys236 f Cl ink to Asp111 f R4.
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The above figures are
reproduced from the cited reference
with permission from Elsevier
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Secondary reference #14
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Title
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Interactions of phosphate ligands with escherichia coli aspartate carbamoyltransferase in the crystalline state.
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Authors
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R.B.Honzatko,
W.N.Lipscomb.
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Ref.
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J Mol Biol, 1982,
160,
265-286.
[DOI no: ]
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PubMed id
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Figure 1.
FIG. 1. ic drawing f egulator~~oatal?;tic unit of aspartate carbano?-ltranf'ernse. hobvirlg
vir down the ole~wlar -fold axis rom the outside f he molrwle. Helicrs are shown as ylindrrs
nd strands of p-stucture as arrows. The phosphate crevice is he cleft between he 2 omains of the-
atalytic chain. Arginine residues 54. 05 and 67. which ind hosphate ligands. comr from helix HZ.
trand 84 and heli H6, respectively. The allosteric site for nucleotidrs is in a pocket of the
allosteric ffector domain. Residues that ind he base f wlroside triphosphatw riginate rom he S-
t,rrminal trand, and residues that are ear thr triphosphatc estw moiety rr a part of thr loo;)
crmnrcting strands S2' o X3'.
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The above figure is
reproduced from the cited reference
with permission from Elsevier
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Secondary reference #15
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Title
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Interactions of metal-Nucleotide complexes with aspartate carbamoyltransferase in the crystalline state.
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Authors
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R.B.Honzatko,
W.N.Lipscomb.
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Ref.
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Proc Natl Acad Sci U S A, 1982,
79,
7171-7174.
[DOI no: ]
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PubMed id
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Secondary reference #16
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Title
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Gross quaternary changes in aspartate carbamoyltransferase are induced by the binding of n-(Phosphonacetyl)-L-Aspartate: a 3.5-A resolution study.
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Authors
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J.E.Ladner,
J.P.Kitchell,
R.B.Honzatko,
H.M.Ke,
K.W.Volz,
A.J.Kalb,
R.C.Ladner,
W.N.Lipscomb.
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Ref.
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Proc Natl Acad Sci U S A, 1982,
79,
3125-3128.
[DOI no: ]
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PubMed id
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Secondary reference #17
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Title
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A 3.0-A resolution study of nucleotide complexes with aspartate carbamoyltransferase.
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Authors
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R.B.Honzatko,
H.L.Monaco,
W.N.Lipscomb.
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Ref.
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Proc Natl Acad Sci U S A, 1979,
76,
5105-5109.
[DOI no: ]
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PubMed id
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Secondary reference #18
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Title
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Three-Dimensional structures of aspartate carbamoyltransferase from escherichia coli and of its complex with cytidine triphosphate.
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Authors
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H.L.Monaco,
J.L.Crawford,
W.N.Lipscomb.
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Ref.
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Proc Natl Acad Sci U S A, 1978,
75,
5276-5280.
[DOI no: ]
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PubMed id
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Secondary reference #19
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Title
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Binding site at 5.5 angstroms resolution of cytidine triphosphate, The allosteric inhibitor of aspartate transcarbamylase from escherichia coli. Relation to mechanisms of control
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Authors
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W.N.Lipscomb,
B.F.P.Edwards,
D.R.Evans,
S.C.Pastra-Landis.
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Ref.
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structure and conformation ...
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Secondary reference #20
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Title
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Aspartate transcarbamoylase from escherichia coli: electron density at 5.5 a resolution.
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Authors
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S.G.Warren,
B.F.Edwards,
D.R.Evans,
D.C.Wiley,
W.N.Lipscomb.
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Ref.
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Proc Natl Acad Sci U S A, 1973,
70,
1117-1121.
[DOI no: ]
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PubMed id
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