ABSTRACT The method of partial least squares projections to latent structures (PLS) has been used... more ABSTRACT The method of partial least squares projections to latent structures (PLS) has been used to derive quantitative relationships between measured reduction potentials and structures of some Pseudomonas aeruginosa azurin mutants with changes in position 121. The models indicate that relatively few properties of the amino acids explain the experimental results obtained. Electronic characteristics exert a major influence on the models but also factors associated with acid-base strength, lipophilicity as well as with hydrogen bonding have large impact. The calculations also show that no single amino acid, natural or synthetic, can meet all of the optimum requirements of position 121 but indicate that a fine balance between various factors is a necessary prerequisite for a proper choice of residue to obtain a mutant with the desired properties.
The method of partial least squares projections to latent structures (PLS) has been used to deriv... more The method of partial least squares projections to latent structures (PLS) has been used to derive quantitative relationships between measured reduction potentials and structures of some Pseudomonas aeruginosa azurin mutants with changes in position 121. The models indicate that relatively few properties of the amino acids explain the experimental results obtained. Electronic characteristics exert a major influence on the models but also factors associated with acid-base strength, lipophilicity as well as with hydrogen bonding have large impact. The calculations also show that no single amino acid, natural or synthetic, can meet all of the optimum requirements of position 121 but indicate that a fine balance between various factors is a necessary prerequisite for a proper choice of residue to obtain a mutant with the desired properties.
We previously selected four strains of Saccharomyces cerevisiae for their ability to produce the ... more We previously selected four strains of Saccharomyces cerevisiae for their ability to produce the aquaporin Fps1 in sufficient yield for further study. Yields from the yeast strains spt3Δ, srb5Δ, gcn5Δ and yTHCBMS1 (supplemented with 0.5 μg/mL doxycycline) that had been transformed with an expression plasmid containing 249 base pairs of 5' untranslated region (UTR) in addition to the primary FPS1 open reading frame (ORF) were 10-80 times higher than yields from wild-type cells expressing the same plasmid. One of the strains increased recombinant yields of the G protein-coupled receptor adenosine receptor 2a (A2aR) and soluble green fluorescent protein (GFP). The specific molecular mechanisms underpinning a high-yielding Fps1 phenotype remained incompletely described. Polysome profiling experiments were used to analyze the translational state of spt3Δ, srb5Δ, gcn5Δ and yTHCBMS1 (supplemented with 0.5 μg/mL doxycycline); all but gcn5Δ were found to exhibit a clear block in translat...
ABSTRACT The method of partial least squares projections to latent structures (PLS) has been used... more ABSTRACT The method of partial least squares projections to latent structures (PLS) has been used to derive quantitative relationships between measured reduction potentials and structures of some Pseudomonas aeruginosa azurin mutants with changes in position 121. The models indicate that relatively few properties of the amino acids explain the experimental results obtained. Electronic characteristics exert a major influence on the models but also factors associated with acid-base strength, lipophilicity as well as with hydrogen bonding have large impact. The calculations also show that no single amino acid, natural or synthetic, can meet all of the optimum requirements of position 121 but indicate that a fine balance between various factors is a necessary prerequisite for a proper choice of residue to obtain a mutant with the desired properties.
The method of partial least squares projections to latent structures (PLS) has been used to deriv... more The method of partial least squares projections to latent structures (PLS) has been used to derive quantitative relationships between measured reduction potentials and structures of some Pseudomonas aeruginosa azurin mutants with changes in position 121. The models indicate that relatively few properties of the amino acids explain the experimental results obtained. Electronic characteristics exert a major influence on the models but also factors associated with acid-base strength, lipophilicity as well as with hydrogen bonding have large impact. The calculations also show that no single amino acid, natural or synthetic, can meet all of the optimum requirements of position 121 but indicate that a fine balance between various factors is a necessary prerequisite for a proper choice of residue to obtain a mutant with the desired properties.
We previously selected four strains of Saccharomyces cerevisiae for their ability to produce the ... more We previously selected four strains of Saccharomyces cerevisiae for their ability to produce the aquaporin Fps1 in sufficient yield for further study. Yields from the yeast strains spt3Δ, srb5Δ, gcn5Δ and yTHCBMS1 (supplemented with 0.5 μg/mL doxycycline) that had been transformed with an expression plasmid containing 249 base pairs of 5' untranslated region (UTR) in addition to the primary FPS1 open reading frame (ORF) were 10-80 times higher than yields from wild-type cells expressing the same plasmid. One of the strains increased recombinant yields of the G protein-coupled receptor adenosine receptor 2a (A2aR) and soluble green fluorescent protein (GFP). The specific molecular mechanisms underpinning a high-yielding Fps1 phenotype remained incompletely described. Polysome profiling experiments were used to analyze the translational state of spt3Δ, srb5Δ, gcn5Δ and yTHCBMS1 (supplemented with 0.5 μg/mL doxycycline); all but gcn5Δ were found to exhibit a clear block in translat...
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