A pharmaceutical company wants to get a particular protein to perform multiple tests with therapeut

This is for the delivery of the third year of bioinformatics biotechnology degree at UAB. It was proposed to do the next job. The realization of this has been between Sancho and Luis Revilla me.
Written Statement Information about protein structure databases general features secondary post-translational modifications and interactions with other protein ligands Predictivament General characteristics secondary structure domains post-translational modification patterns Multialineament sand filter Protocol for cloning Design Selection of first vector selection targets Process improvements structural separation and purification
A pharmaceutical company wants to get a particular protein to perform multiple tests with therapeutic application. This company sand filter has commissioned us to perform a bioinformatic study of the properties of the protein and design a protocol for obtaining, using the tools described in the course of Bioinformatics.
The initial sand filter information for our study is the protein sequence that corresponds to the code that will give the teacher. (UniprotKB Code: P00374) Closing sand filter Reference: Find as much information about the problem in protein databases. Make predictions on physico-chemical properties, structural elements and motifs and domains, interactions with other proteins ... Search multialineaments counterparts and perform with other protein sequences from the same family. Determine whether the three-dimensional structure has been solved and if it matches your prediction. Designing a protocol for cloning, expression and purification of the protein problem: Designing primers (first) to amplify by PCR and cloned the protein sand filter in a plasmid vector (pET28a, pET41a, pET21a). Determine what will be the strategy to make the process of purification of the protein based on the characteristics of the plasmid.
In the database of UniProt can be observed that corresponds sand filter to the protein dihydrofolate reductase. Is a key enzyme involved in the metabolism so that the de novo biosynthesis of mitochondria timidilat. Catalyzes a reaction essential for the synthesis of glycine and purines. Is an enzyme that reduces the acid in dihydrofolate tetrahidrofolat using the NAPH as an electron donor. Tetrahidrofolat this is a basic component for the synthesis of these purines, thymidine monophosphate and certain amino acids.
If you do a search on Brenda confirms that its catalytic activity is defined by the KM for NADPH was 4.0 mM and 2.7 mM is the dihydrofolate. PH range where it works best is between 4.2 and 5.2.
As indicated genecards has a molecular mass of approximately 21,500 Da, corresponding as seen in the PDB that has 187 amino acids, in addition to post-translational modifications.
As can be seen in terms of secondary structure has 22% of α helix and 32% β sheet, according to the structural classification is a protein means that class Alpha / Beta. In this case 4DDR used for all the work with reference to data from the PDB because it presents only the union of NADPH and another product, but we believe that its structure sand filter and data is significant enough as to be used, since the data were taken with a high resolution X-ray With a maximum data resolution and minimum 2.05 2.12 angstrom resolution.
The protein has no disulfide bond, which would be impossible since it only has a single cysteine. Regarding the post-translational modifications include acetylation is a two phosphorylation and ubiquitination, as can be seen on the website PhosphoSitePlus. These changes are minimal sand filter and the protein remains functional although not occur, although the kinetics changed. Finally, the initial methionine is removed.
On the left column indicates the atom interacts NADPH and the bottom row shows the amino acid involved. There are three types of joints present: hydrogen (pink), unions sand filter hydrophobic (green) and hydrogen laid mediated by water (blue).
Has some connection with the cyclin A2 and should therefore play a role in regulating the cell cycle and in the maintenance of chromosomes as seen interacts with timidilat synthase, which is closely related, since as s' said participating in the same metabolic pathway.
Since it is a human protein that in humans is we want to provide better rely on data referring to organisms sand filter evolutionarily close as was possible and other mammals sand filter in vivo, as in yeast. By the vectors used, probably used as a microorganism Escherichia coli.
This sequence homology is quite