Purification of near identical Escherichia coli aspartate transcarbamoylase holoenzyme hybrids by electroelution method
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چکیده
منابع مشابه
Thermodynamics of assembly of Escherichia coli aspartate transcarbamoylase.
Reaction microcalorimetry and potentiometry have been used to define the thermodynamics of assembly of Escherichia coli aspartate transcarbamoylase (aspartate carbamoyltransferase, carbamoylphosphate:L-aspartate carbamoyltransferase, EC 2.1.3.2) from its catalytic and regulatory subunits and the linkage between assembly and proton binding. Over the pH range 7-9.5 and the temperature range 15-30...
متن کاملAssembly of catalytic subunits of aspartate transcarbamoylase from Escherichia coli.
participate in different ways in the assembly of the complex. The positioning of a subunit in the complex may be crucial to its function. The dynamic equilibrium between various assembly states of a heteropolymer cannot be treated as if the assembly involves structurally homogeneous elements whose association reactions can be described by a single interaction constant. On the contrary, each sta...
متن کاملPurification of aspartate transcarbamoylase from Pseudomonas syringae.
The aspartate transcarbamoylase (ATCase) from Pseudomonas syringae has been purified. The purified enzyme was shown by SDS-PAGE to give two bands. Unambiguous results from N-terminal sequencing suggested that each band represented a homogeneous polypeptide. The M(r) (relative molecular mass) of the polypeptides was estimated to be 47 kDa and 34 kDa. The M(r) of the holoenzyme determined by gel ...
متن کاملStabilization of the R allosteric structure of Escherichia coli aspartate transcarbamoylase by disulfide bond formation.
Here we report the first use of disulfide bond formation to stabilize the R allosteric structure of Escherichia coli aspartate transcarbamoylase. In the R allosteric state, residues in the 240s loop from two catalytic chains of different subunits are close together, whereas in the T allosteric state they are far apart. By substitution of Ala-241 in the 240s loop of the catalytic chain with cyst...
متن کاملEffectors of Escherichia coli aspartate transcarbamoylase differentially perturb aspartate binding rather than the T-R transition.
New systematic methods developed for equilibrium isotope exchange kinetics have been used to analyze the effects of activator ATP and inhibitor CTP with Escherichia coli aspartate transcarbamoylase. This indepth approach requires (a) variation of [modifier] with fixed subsaturating levels of substrates, and (b) variation of at least three combinations of reactant-product pairs in constant ratio...
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ژورنال
عنوان ژورنال: SEIBUTSU BUTSURI KAGAKU
سال: 2004
ISSN: 0031-9082,1349-9785
DOI: 10.2198/sbk.48.67