Protein stability determinants

Zoecklein B.W. (1988c). Protein stability determination in juice and wine. Virginia Cooperative Extension, Publication No. 463-015. 

GPCR function has been shown to be regulated by several different mechanisms. The number of receptors on the plasma membrane may be regulated by transcription, mRNA stability, biosynthetic processing, and protein stability. In addition, the function of receptors in the plasma membrane can be influenced by regulatory phosphorylation and by association with other proteins that determine the subcellular location of receptors relative to other signaling molecules. 

Electrostatic stabilization, 181, 195,225-228 Empirical valence bond model, see Valence bond model, empirical Energy minimization methods, 114-117 computer programs for, 128-132 convergence of, 115 local vr. overall minima, 116-117 use in protein structure determination,... 

Enzymatic reactions are influenced by a variety of solution conditions that must be well controlled in HTS assays. Buffer components, pH, ionic strength, solvent polarity, viscosity, and temperature can all influence the initial velocity and the interactions of enzymes with substrate and inhibitor molecules. Space does not permit a comprehensive discussion of these factors, but a more detailed presentation can be found in the text by Copeland (2000). Here we simply make the recommendation that all of these solution conditions be optimized in the course of assay development. It is worth noting that there can be differences in optimal conditions for enzyme stability and enzyme activity. For example, the initial velocity may be greatest at 37°C and pH 5.0, but one may find that the enzyme denatures during the course of the assay time under these conditions. In situations like this one must experimentally determine the best compromise between reaction rate and protein stability. Again, a more detailed discussion of this issue, and methods for diagnosing enzyme denaturation during reaction can be found in Copeland (2000). 

In order to make molecular farming commercially profitable, recombinant proteins must be produced at a sufficiently high yield and in an active form. It has become clear that, for high-level protein accumulation, the stability of transgene expression can be as important as the expression level itself. The quantity of protein is determined by the rate of protein synthesis, assembly as well as proteolytic degradation [83]. 

One of the most important considerations for the improvement of protein yields is subcellular protein targeting, because the compartment in which a recombinant protein accumulates strongly influences the interrelated processes of folding, assembly and post-translational modification. All of these contribute to protein stability and hence help to determine the final yield [88]. 

CD spectra, particularly in the near-UV (see Support Protocol I) reflect the dynamics of the chromophore and may therefore show dependence on temperature. It is important to stabilize the temperature reproducibly. Accurate temperature control is particularly important in denaturation experiments to determine protein stability. 

Ultrasonic methods infrared scanning for emulsion stability determination, 597-598 spectrometry, emulsion droplet size determination, 581 velocimetry, to measure fat, 572 Ultraviolet (UV). see also Spectrophotometry protein analysis, CD, 219-243 protein concentrations by... 

Midgley, C.A. and Lane, D.P. (1997) P53 protein stability in tumor cells is not determined by mutation but is dependent on MDM2 binding. Oncogene, 15, 1179-1189. 

Replication starts by the separation of the strands of DNA and the formation of a local bubble at a specific DNA site called the origin of replication (ori). A helicase enzyme uses energy from ATP hydrolysis to effect this action. Single-strand DNA binding proteins stabilize the strands during the subsequent steps. The original DNA strands will function as the templates that will direct synthesis of the complementary strands. A nucleotide on the template strand will determine which deoxyribonucleotide (dNTP) will be incorporated in the newly synthesized strand. This replica-... 

The proteins to be included in VLPs should be those necessary to confer the desired degree of immunogenicity. As a consequence, VLPs are often composed of more than one protein. In order to use baculovirus-infected insect cells to produce VLPs it is necessary to predefine the proteins to be included, since the expression of these proteins could determine the stability (Hyatt et al., 1993) and location - intra- or extracellular - of the particle (French and Roy, 1990). 

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