Background: You are working in a biochemistry/molecular biology lab that special
ID: 146075 • Letter: B
Question
Background: You are working in a biochemistry/molecular biology lab that specializes in protein applications (purification, identification, etc). The lab has been working with a new organism This organism is single-cellular and eukaryotic and appears to be related to yeast. You are entering the lab and have been assigned characterizing this new organism. Proteins and Proteomics 1. How would you begin purifying and separating the proteome of the organism? What are properties of proteins that we can take advantage of for this purpose? 2. How would you determine if there are similar proteins in this organism compared to yeast western blot technique) 3. How would you determine what proteins interact together? 4. How much you determine the amino acid sequence of a protein? 5. How would knowing the amino acid sequence of a protein help you for nucleic acids?Explanation / Answer
1. Proteins are purified by enzyme protesaes which digest the protein. DNAase and RNAase are used for removing DNA and RNA from the cells. Western blot is used to separate the proteins.
Proteins are separated by various methods like SDS page which used the molecular weight present on the proteins to separate them, centrifugation which used different molecular weight of the protein, Hydrophobic chromatography is used for separation of proteins according to their hydrophobic property.
2. We can find out similar proteins in yeast using NCBI blast.
3. We can find the interaction of proteins using wet lab methods like yeast 2 hybrid method, co-immunoprecipitaion. Using bioinformatics approaches like Using STRING database.
4. By Sanger sequencing, or NGS, next generation sequencing we can seduce the sequence of protein.
5. By knowing the amino acid sequence , we can deduce the DNA of that protein using server like protparam server.
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