Test if gluten has been metabolized.

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This page is for Steven's research.

http://eprints.uniss.it/5145/1/Forteschi_M_Study_of_peptidases_involved.pdf 33-mer peptide: LQLQPFPQPQLPYPQPQLPYPQPQLPYPQPQPF


http://www.sciencedirect.com/science/article/pii/0009898190900824 Detection and estimation of the barley prolamin content of beer and malt to assess their suitability for patients with coeliac disease -a study confirming that beer contains levels of gluten antigens harmful to Celiacs.

http://www.sciencemag.org/content/297/5590/2275.full Structural Basis for Gluten Intolerance in Celiac Sprue -The antigenicity of gluten seems to be due to proline and glutamine rich peptides, the most prevalent one being the 33-mer listed above. The sequence seems to be fairly conserved across barley and wheat.Three distinct patient-specific T cell epitopes identified previously in T cell proliferation assays are present in this peptide, namely, PFPQPQLPY, PQPQLPYPQ (three copies), and PYPQPQLPY (two copies).

http://www.sunrisescience.com/Files/pTEF-MF.pdf pTEF-MF constitutive yeast vector marked for secretion -MF-alpha secretion tag for yeast is used in this plasmid. I used the plasmid map to get the tag's sequence and BLASTed it. A 100% match confirmed that the sequence is present in yeast. We will attach this to the N-terminus of Kumamolisin, hopefully enabling the protein to be secreted in yeast.

http://www.jbc.org/content/263/13/6209.full.pdf Prepro-a-factor Has a Cleavable Signal Sequence -MF-alpha signal sequence is attached to the animo-terminus of MF-alpha and cleaved in the golgi.

http://synbio.org.uk/dna-assembly/guidetogibsonassembly.html Gibson assembly

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC391546/pdf/pnas00616-0033.pdf Alphafactor- directed synthesis and secretion of mature foreign proteins in Saccharomyces cerevisiae

http://www.springerlink.com/content/l2115622t21t728w/fulltext.pdf Engineering of protein secretion in yeast: strategies and impact on protein production

http://ajpgi.physiology.org/content/291/4/G621.long Highly efficient gluten degradation with a newly identified prolyl endoprotease: implications for celiac disease

http://onlinelibrary.wiley.com/doi/10.1002/jms.1667/full Identification of N-glycosylation in prolyl endoprotease from Aspergillus niger and evaluation of the enzyme for its possible application in proteomics

http://mcl1.ncifcrf.gov/wlo_pubs/261.pdf enzyme structure and kinetics for Kumamolisin