You are sequencing a protein. Previous work indicates that thisprotein consists of two polypeptide chains that are held togetherby a single disulfide bond. You perform two experiments and get theresults indicated below. From this âexperimentalâ information.Determine the sequences of the two polypeptide chains and explainhow you obtain this?
Experiment #1
In this experiment, you cleave the disulfide bond with performicacid, separate the two chains, and sequence them by treatment withthe proteolytic enzymes trypsin and chymotrypsin, followed by Edmandegradation to determine the sequences of the each of the resultingfragments. Through using an overlap-peptide type of method, youshould be able to determine the original sequence of the twoseparate polypeptide chains
Trypsin Treatment
chain 1:
I
LK
NQR
HSFWK
FWPCDR
chain 2:
GAR
MPAK
LSYSK
LSCFGR
LAACNIYF
Chymotrpsin Treatment
chain 1:
W
W
HSF
PCDRI
KNQRLKF
chain 2:
F
GRGARLSY
MPAKLSCF
SKLAACNIY
You are sequencing a protein. Previous work indicates that thisprotein consists of two polypeptide chains that are held togetherby a single disulfide bond. You perform two experiments and get theresults indicated below. From this âexperimentalâ information.Determine the sequences of the two polypeptide chains and explainhow you obtain this?
Experiment #1
In this experiment, you cleave the disulfide bond with performicacid, separate the two chains, and sequence them by treatment withthe proteolytic enzymes trypsin and chymotrypsin, followed by Edmandegradation to determine the sequences of the each of the resultingfragments. Through using an overlap-peptide type of method, youshould be able to determine the original sequence of the twoseparate polypeptide chains
Trypsin Treatment
chain 1:
I
LK
NQR
HSFWK
FWPCDR
chain 2:
GAR
MPAK
LSYSK
LSCFGR
LAACNIYF
Chymotrpsin Treatment
chain 1:
W
W
HSF
PCDRI
KNQRLKF
chain 2:
F
GRGARLSY
MPAKLSCF
SKLAACNIY
