Two-hybrid

McCormack D A, Kroes G J and Neuhauser D 1998 Resonance affected scattering Comparison of two hybrid methods involving filter diagonalization and the Lanczos method J. Chem. Phys. 109 5177... 

The Aromax process was developed in the early 1970s by Toray Industries, Inc. in Japan (95—98). The adsorption column consists of a horizontal series of independent chambers containing fixed beds of adsorbent. Instead of a rotary valve, a sequence of specially designed on—off valves under computer control is used to move inlet and withdrawal ports around the bed. Adsorption is carried out in the Hquid phase at 140°C, 785—980 kPA, and 5—13 L/h. PX yields per pass is reported to exceed 90% with a typical purity of 99.5%. The first Aromax unit was installed at Toray s Kawasaki plant in March 1973. In 1994, IFP introduced the Eluxyl adsorption process (59,99). The proprietary adsorbent used is designated SPX 3000. Individual on-off valves controlled by a microprocessor are used. Raman spectroscopy to used to measure concentration profiles in the column. A 10,000 t/yr demonstration plant was started and successfully operated at Chevron s Pascagoula plant from 1995—96. IFP has Hcensed two hybrid units. 

All of the predicted structures are at least reasonably good. The two hybrid functionals produce the best structures, in excellent agreement with the experimental geometry. The SVWN and SVWNS functionals both produce good structures, while the BLYP geometry is the least accurate. 

The Two-Hybrid System to Identify Proteins Involved in Specific Protein-Protein Interactions... 

Chien, C.-X, et al., 1991. The two-hybrid system A method to identify and clone genes for proteins that interact with a protein of interest. Proceedings of the National Academy of Sciences U.S.A. 88 9578 — 9582. 

It may be proper at this stage to lead the reader back to the stage where we constructed the localized orbitals of a CH2 group. At that time two valence orbitals were set aside—the 2pv orbital, and the outer (2s, 2pr) hybrid. Both of these orbitals lie in the. r, y plane. Now in our description of cyclopropane, we used bond orbitals to describe the CC bonding these bond orbitals are derived from in-plane (xy y) hybrids on each carbon. The two hybrids which are required on each carbon atom—in ordet to participate in two bond orbitals—are built precisely from the 2py orbital and the (2s, 2pj.) out combination on each CH2 group. 

Biological raw data are stored in public databanks (such as Genbank or EMBL for primary DNA sequences). The data can be submitted and accessed via the World Wide Web. Protein sequence databanks like trEMBL provide the most likely translation of all coding sequences in the EMBL databank. Sequence data are prominent, but also other data are stored, e.g.yeast two-hybrid screens, expression arrays, systematic gene-knock-out experiments, and metabolic pathways. 

PIAS (protein inhibitors of activated STATs) proteins were first discovered in yeast-two-hybrid screens as interacting molecules with STAT transcription factors. The mammalian family consists ofthe founding member PIAS3, which was described as a repressor of STAT3, and three additional members, PIAS1, PIASy (also known as PIAS4), and PIASx (also known as... 

A linear arrangement of electron pairs requires two hybrid orbitals, and so we mix an s-orbital with a /7-orbital to obtain two s/7-hybrid orbitals ... 

Noting that the bond angle of an sp hybridized atom is 109.5° and that of an sp1 hybridized atom is 120°, do you expect the bond angle between two hybrid orbitals to increase or decrease as the s-character of the hybrids is increased ... 

Given the information in Exercise 3.47, plot a graph showing how the hybridization depends on the angle between two hybrids formed from an s-orbital and two p-orbitals, and confirm that it ranges from 90° when no s-orbital is included in the mixture to 120° when the hybridization is sp2. 

Waters PJ, Scriver CR, Parniak MA Homomeric and heteromeric interactions between wild-type and mutant phenylalanine hydroxylase subunits evaluation of two-hybrid approaches for functional analysis of mutations causing hyperphenylalanine-mia. Mol Genet Metab 2001 73 230. 

Both inner atoms have steric numbers of 4 and tetrahedral electron group geometry, so both can be described using s p hybrid orbitals. All four hydrogen atoms occupy outer positions, and these form bonds to the inner atoms through 1 s-s p overlap. The oxygen atom has two lone pairs, one in each of the two hybrid orbitals not used to form O—H bonds. 

A pair of sp hybrid orbitais forms by interacting an s atomic orbital and one p atomic orbital. The two hybrids point in opposite directions. 

An sp-hybridized atom has two hybrid orbitals separated by 180°. The remaining two atomic p orbitals are perpendicular to the hybrids and perpendicular to each other. 

The double bond in ethylene contains one a bond and one 7r bond. The a bond forms from the end-on overlap of two hybrid orbitals, and the 7i bond forms from the side-by-side overlap of two atomic p orbitals. Figure 10-21 shows the complete orbital picture of the bonding in ethylene. Ethylene is the simplest of a class of molecules, the alkenes, all of which contain CDC double bonds. The alkenes are the subject of our Box on page 404. 

Figure 5.1. Yeast two-hybrid system. Interaction of proteins X and Y upstream of a reporter gene leads to transcriptional activation. Protein X is part of a fusion protein that binds to a site on DNA upstream of the reporter gene by means of a DNA binding domain. Protein Y is part of a fusion protein that contains a transcriptional activation domain. Interaction of proteins X and Y places the activation domain in the vicinity of the reporter gene and stimulates its transcription.

A major challenge in the use of two-hybrid systems is the elimination of false positives. These clones result from activation of reporter... 

Figure 5.2. High-throughput mating assay for two-hybrid protein interaction screening. Yeast strains containing individual bait and prey clones are combined in a well and allowed to mate. Diploids are then selected and scored for a protein-protein interaction using the selection provided by the transcriptional reporter gene.

Figure 5.3. Systematic mating ofyeast two-hybrid bait and prey pools. Each yeast ORF was cloned individually into both as a DNA binding domain fusion (bait) and activation domain fusion (prey). The bait fusions were introduced into a MATa strain and the prey fusions were introduced into a MATa strain. The bait and prey fusions were pooled in sets of 96 clones to generate a total of 62 pools of each. The pools were systematically mated (62 x 62) in a total of 3844 crosses. Interacting clones were selected and the bait and prey inserts were PCR amplified and sequenced to determine their identify. Figure adapted from Ito et al. (2001).

---