BIOCHEMISTRY PROBLEM:
I have a sample that contains three small peptides (intact sample run on lane 2 of SDS-PAGE gel). Theseproteins were separated from one another through column chromatography and an attempt at sequencingwas made. The researchers were able to get through the entire sequencing of peptide 1. Here is thesequence: SYSQEHCDNFEGWCDHFVVTYGWSERACDI. Peptides two and three were not sequenced, but theproteolytic digests were completed (information below).
To complete:
(a) Show the full three-dimensional structure of the 5-amino acid fragment from the digest of peptide 1 with V8 protease.
(b) Deduce and show the sequence of peptides 2 and 3
(c) Show the outcome, by diagraming the progress on an SDS-PAGE gel, of the following separatory method(the first step is mandatory, the second step is one you have to make choices!)
First, the peptides were applied to a gel filtration column. A sample from the collection of proteins found inearly fractions were run in column 3 of the gel, and late fractions in column 4. SHOW THE BANDS THATWOULD RESULT ON THE SDS-PAGE GEL.
Next, the fraction (from part one above) containing two peptides was applied to an ion exchange column. The early fractions (flow through) were combined, and a sample was run in column 5 of the SDS-PAGE gel and thelate fractions (after salt wash) were run in column 6. CHOOSE THE TYPE OF RESIN YOU WILL USE, THE pH YOU WILL WORK, THE CALCULATIONS NECESSARY TO MAKE AN APPROPRIATE 200 mM BUFFER AND THEN SHOW THE BANDS THAT WOULD RESULT ON THE SDS-PAGE GEL.
Note: the standard set run in lane one on the SDS-PAGE gel contains the proteins with the following masses(all KDa): 1, 3.5, 6.5, 14.2, 17, 26.5.
Here is information on the last two peptides that I separated â
A pure sample of peptide 2 gives the following cleavage pattern when cleaved with chymotrypsin: L, SY, HW, SMEHF, GKPVY. This same peptide cleaved with thermolysin gives the following fragments: VY, SYS, MEHF, LHWGKP
A pure sample of peptide 3 gives the following cleavage pattern when cleaved with trypsin: FGW, WQLR, GHIFR, AVYDMK. Peptide 3 cleaved with thermolysin gives the following fragments: A, VYD, MKGH, IFRWQ, LRFGW.
**I have already found the sequence for peptide 2 and 3 I think and know what the SDS page will look like** I just need help figuring out the rest of the blanks in this sheet:
Thank you in advance: