This question has been posted previously with parts A B and C already answered. Please focus on only D, E and F Provided is the question alongside with the added materials next to it. Thank you 2. You are in a South American rain forest looking for naturally occuring peptides with potential as drugs.You have a mobile biochemistry lab with common reagents and enzymes, an amino-acid analyzer, gel-filtration and ion-exchange chromatography, and electrophoresis. You also have an Edman Sequenator, butyou have contaminated one or more of your reagents, and as a result, you cannot sequence peptideslonger than about 12 residues before contaminants obscure the results. While screening extracts from theovaries of an tropical orchid, you find a peptide with potential as an antiviral. Deduce its amino-acidsequence using the available tools.1) MW by electrophoresis can tell you how big a sequencing problem you are up against.Result: about 40002) Amino-acid analysis can help you decide how to fragment the peptide for sequencing:Result: A2C2D2E4FG3HKLMN2P2Q2R4S4T3W3) How many peptides expected from each of these possible cleavage reagents?●• Cyanogen bromide (C-side of M).• Staph. aureus V8 protease (C-side of D and E).• Trypsin (C-side of K and R).4) Cleavage by trypsin followed by gel-filtration chromatography gives the expected 6 products, which yousequence (shown in order of emergence from column):T-1 ETMESSAGEFGR T-2 SQTWALDHSECR T-3 GPQDNKT-4 TCR T-5 NP T-6 R5) Cleavage by Staph. aureus V8 protease followed by gel-filtration chromatography gives theexpected 7 products, which you sequence (shown in order of emergence from column):S-1 RSQTWALD S-2 FGRGPQD S-3 NKTCRNP S-4 SSAGES-5 TME S-6 CRE S-7 HSEThink About It:A. Deduce the primary structure of this polypeptide.B. Why would cyanogen bromide not be a good choice as a cleavage reagent?C. Can you account for the order of elution of trypsin digest peptides from gel-filtration chromatography?D. Predict the order of elution of the tryptic peptides from a cation-exchange column eluted with pH-8.5buffer and a salt gradient.E. Predict the order of elution of the V8 protease peptides from an anion exchange chromatographycolumn eluted with a pH-6.5 buffer and a salt gradient.F. For both sets of peptides, predict the order of elution from a hydrophobic interaction chromatographycolumn, given that the hydrophobic amino acids are V,L,I,F,A, and M. Protein Sequencing ProblemsTo get you started, here is a table with the specificities for some of the enzymaticand chemical procedures used in sequencing proteins:ProcedureEdman DegradationCarboxypeptidase ACarboxypeptidase BCyanogen BromideaminoethylationTrypsinChymotrypsinStaphylococccus aureus V8ProteaseAnhydrous Cyanogen Bromide C side of RSubmaxillarus ProteaseEndoproteinase Lys CAsp-N-proteaseThermolysinPepsinAcid PhosphataseAcid HydrolysisSiteR₁-1R₂C side of N terminus R = any amino acidN side of C terminus R, # Arg, Lys, ProR₁-1 ProN side of C terminus R Arg, Lys, AECysR₁-1 # ProR = MetC side of R₁Cys residueC side of RCside of RC side of RC side of R₁C side of RN side of RN side of R₁N side of RSpecificityC side of R₁Breaks all peptidebondsRn+1R = Met, TrpReacts with Cys to giveAECysR = Lys, Arg, AECysR₂+1 #ProR = Phe, Trp, Tyr, LeuR₁+1 #ProR = Asp D, Glu ER = ArgR = Lys KR = Asp D, Glu ECommentR=Leu, Asp, Glu, Phe,Tyr, TrpR₁+1 # ProR = Asp, GluIneffective for blocked N terminiRemoves 1 (usually up to 4) residuessequentiallyRemoves 1 (usually up to 4) residuessequentiallyHighly SpecificHighly SpecificHighly SpecificSometimes cleaves at other sitesR = Leu, Ile, Phe, Trp, Tyr, Sometimes cleaves at other sitesValR₁+1 #ProThe table uses terms like "R". I've drawn a short piece of polypeptide so youcan visualize where these cleavages are taking place.N side of R₁Fairly nonspecificHighly SpecificC side of R₁If the amino acids are listed with dashes connecting them, they are in sequence.If the amino acids are listed separated by commas, they are not in order.For example.....If I treat G-K-A-V-M with trypsin I will get two pieces. One will be (G-K) andthe other will be (A-V-M). If I only know the composition (and not the actualsequence for these pieces) it would be written (G, K) and (A, M, V).

Question

This question has been posted previously with parts A B and C already answered. Please focus on only D, E and F Provided is the question alongside with the added materials next to it. Thank you 2. You are in a South American rain forest looking for naturally occuring peptides with potential as drugs.You have a mobile biochemistry lab with common reagents and enzymes, an amino-acid analyzer, gel-filtration and ion-exchange chromatography, and electrophoresis. You also have an Edman Sequenator, butyou have contaminated one or more of your reagents, and as a result, you cannot sequence peptideslonger than about 12 residues before contaminants obscure the results. While screening extracts from theovaries of an tropical orchid, you find a peptide with potential as an antiviral. Deduce its amino-acidsequence using the available tools.1) MW by electrophoresis can tell you how big a sequencing problem you are up against.Result: about 40002) Amino-acid analysis can help you decide how to fragment the peptide for sequencing:Result: A2C2D2E4FG3HKLMN2P2Q2R4S4T3W3) How many peptides expected from each of these possible cleavage reagents?●• Cyanogen bromide (C-side of M).• Staph. aureus V8 protease (C-side of D and E).• Trypsin (C-side of K and R).4) Cleavage by trypsin followed by gel-filtration chromatography gives the expected 6 products, which yousequence (shown in order of emergence from column):T-1 ETMESSAGEFGR T-2 SQTWALDHSECR T-3 GPQDNKT-4 TCR T-5 NP T-6 R5) Cleavage by Staph. aureus V8 protease followed by gel-filtration chromatography gives theexpected 7 products, which you sequence (shown in order of emergence from column):S-1 RSQTWALD S-2 FGRGPQD S-3 NKTCRNP S-4 SSAGES-5 TME S-6 CRE S-7 HSEThink About It:A. Deduce the primary structure of this polypeptide.B. Why would cyanogen bromide not be a good choice as a cleavage reagent?C. Can you account for the order of elution of trypsin digest peptides from gel-filtration chromatography?D. Predict the order of elution of the tryptic peptides from a cation-exchange column eluted with pH-8.5buffer and a salt gradient.E. Predict the order of elution of the V8 protease peptides from an anion exchange chromatographycolumn eluted with a pH-6.5 buffer and a salt gradient.F. For both sets of peptides, predict the order of elution from a hydrophobic interaction chromatographycolumn, given that the hydrophobic amino acids are V,L,I,F,A, and M. Protein Sequencing ProblemsTo get you started, here is a table with the specificities for some of the enzymaticand chemical procedures used in sequencing proteins:ProcedureEdman DegradationCarboxypeptidase ACarboxypeptidase BCyanogen BromideaminoethylationTrypsinChymotrypsinStaphylococccus aureus V8ProteaseAnhydrous Cyanogen Bromide C side of RSubmaxillarus ProteaseEndoproteinase Lys CAsp-N-proteaseThermolysinPepsinAcid PhosphataseAcid HydrolysisSiteR₁-1R₂C side of N terminus R = any amino acidN side of C terminus R, # Arg, Lys, ProR₁-1 ProN side of C terminus R Arg, Lys, AECysR₁-1 # ProR = MetC side of R₁Cys residueC side of RCside of RC side of RC side of R₁C side of RN side of RN side of R₁N side of RSpecificityC side of R₁Breaks all peptidebondsRn+1R = Met, TrpReacts with Cys to giveAECysR = Lys, Arg, AECysR₂+1 #ProR = Phe, Trp, Tyr, LeuR₁+1 #ProR = Asp D, Glu ER = ArgR = Lys KR = Asp D, Glu ECommentR=Leu, Asp, Glu, Phe,Tyr, TrpR₁+1 # ProR = Asp, GluIneffective for blocked N terminiRemoves 1 (usually up to 4) residuessequentiallyRemoves 1 (usually up to 4) residuessequentiallyHighly SpecificHighly SpecificHighly SpecificSometimes cleaves at other sitesR = Leu, Ile, Phe, Trp, Tyr, Sometimes cleaves at other sitesValR₁+1 #ProThe table uses terms like &#34;R&#34;. I've drawn a short piece of polypeptide so youcan visualize where these cleavages are taking place.N side of R₁Fairly nonspecificHighly SpecificC side of R₁If the amino acids are listed with dashes connecting them, they are in sequence.If the amino acids are listed separated by commas, they are not in order.For example.....If I treat G-K-A-V-M with trypsin I will get two pieces. One will be (G-K) andthe other will be (A-V-M). If I only know the composition (and not the actualsequence for these pieces) it would be written (G, K) and (A, M, V).

Accepted Answer

Amino acids classified as hydrophobic have side chains (R groups) that have a propensity to resist…