facilitates storage and use of the high energy of the adenine nucleotides, involved in maintenance of equilibrium among adenine nucleotides and maintenance of energy charge, important to energy economy of living systems
facilitates storage and use of the high energy of the adenine nucleotides, involved in maintenance of equilibrium among adenine nucleotides and maintenance of energy charge, important to energy economy of living systems
0.8-1 mol Zn2+ for wild-type and mutants H138N, D153C and D153T, 0.6 mol Zn2+ for mutant D153T, or 0.34 mol Zn2+ for mutant C130H per mol protein, atomic absorption spectrophotometry
optimization of stabilizing interactions connecting distant polypeptide regions, Lys19-Glu202 and Arg116-Glu198 ion pairs are formed in enzyme mutant AKm1, hydrophobic packing is improved by incorporating Tyr109, Val193, and Ile211 into enzyme mutant AKm2
purified recombinant enzyme mutants AKm1 and AKm2 in complex with inhibitor Ap5A, hanging drop vapour diffusion method, mixing of 30 mg/ml AKm1 or 18 mg/ml AKm2 in 10 mM HEPES pH 7.0, and 4 mM Ap5A, with an equal amount of a reservoir solution containing 18% w/v PEG 3350, 100 mM lithium sulfate, and 100 mM Bis-Tris, pH 5.5, for mutant AKm1 and containing 22% w/v PEG 3350, and 200 mM calcium chloride for mutant AKm2, 20°C, X-ray diffraction structure determination and analysis at 2.990 and 1.65 A resolution, respectively
thermostabilization of full-length protein. Cells harboring the mutant fragment pair with concomitant expression of isolated N-terminus, amino acids 1-76 and C-terminus, amino acids 77-217 show weak complementation of Escherichia coli mutant after fusion with polypeptides that strongly associate
modest increase in stability, 3 degrees increase in melting temperature. At temperatures of 20°C to 45°C, 50% loss of activity, with subsequent increase at higher temperatures. rigidification of he overall structure through stabilization of a polypeptide loop containing R199 that is part of the ATP-binding site
no functional complemetation of temperature-sensitive Escherichia coli mutant by concomitant expression of isolated N-terminus, amino acids 1-76 and C-terminus, amino acids 77-217. Weak complementation after fusion with polypeptides that strongly associate
construction of chimeric adenylate kinase mutants designed by local structural entropy optimization and structure-guided mutagenesis. Generation of LSE-optimized AK variant (AKlse, formerly known as AKlse4) by substituting residues from a mesophilic Bacillus subtilis AKmeso with those of psychrophilic Bacillus globisporus and thermophilic Geobacillus stearothermophilus AKs, AKpsycrho and AKthermo, respectively. Using this AKlse as a template, two additional stable AK mutants, AKm1 and AKm2, formerly known as AKlse4m1 and AKlse4m2, respectively, are generated. Mutant crystals structures analysis, detailed overview
construction of chimera between the mesophilic Bacillus subtilis enzyme and the thermophilic Bacillus stearothermophilus enzyme by exchange of one or more of the seven protein segments. Analysis of the chimeras shows spatial separation of stability and activity control with specific contributions of dynamics to catalysis
Structural and mutational analyses of psychrophilic and mesophilic adenylate kinases highlight the role of hydrophobic interactions in protein thermal stability