A New Method for the Isolation of the C-Terminal Peptide of Proteins
187
Table 12.1. Summary of the experimental masses observed in the isolated C-terminal peptide from
the different proteins characterized in this study
Protein
Cytochrome C
IL-2m
TAB-9
Myoglobin
Streptokinase
Mucorpepsin
References
Molecular
Weight (kDA)
11.7
15.6
16.6
17.2
47.3
57.2
(M+HJ'
theor.
1805.8
1522.9
592.3
1983.8
1631.7
1394.6
(M+Hr
1805.9
1522.8
592.2
1984.1
1633.0
1395.7
Sequence
Mal- 92 EDLIAYLK*K* ATNE I04
Ac- 122 WITFAQSIISTLT I34
AC_ 158 AFVTII6'
SUC_140K*DIAK*YK*ELGYQGI53
Suc-403YTGTPIPDNPNDK*415
Ac- 1221 G FAPLASGYENN 134
Bailey JM, Shively JE (1990) Carboxy-terminal sequencing: Formation and hydrolysis of C-terminal
Peptidylthiohydantoins. Biochemistry 29: 3145 - 3156.
Boyd VL, Bozzini M, Zon G, Noble RL, Mattaliano R (1992) Sequencing of peptides and proteins from
the carboxy terminus. Anal. Biochem. 206: 344-352
Butler PJG, Harris JI, Hartley, Bsleberman R (1969) The use of maleic anhydride for the reversible
blocking of amino groups in polypeptide chains. Biochem J 112: 679-689
Gorman JJ, Shiell BJ (1993) Isolation of carboxyl-termini and blocked amino-termini of viral proteins
by high-performance cation-exchange chromatography J. Chromo 646: 193-205.
Isobe T, Ichimura T, Okuyama T (1986) Identification of the C-Terminal Portion of a Protein by Comparative Peptide Mapping. Anal Biochem. 155: 135-140
Kumazaki T, Nakako T, Arisaka F, Ishii S (1986) A novel method for selective isolation of C-terminal
pep tides from tryptic digests of proteins by immobilized anhydrotrypsin: application to structural
analyses of the tail sheath and tube proteins from bacteriophage T4. Proteins 1: 100 -107.
Miller CG, Bailey JM (1995) Biotechnology applications of C-terminal sequence analysis. Genetic
Engineering News 15: 12-16
Murphy CM, Fenselau C (1995) Recognition of Carboxy-Terminal Peptide in Cyanogen Bromide
Digest of Proteins. Anal Chern. 67: 1644-1647
Naughton, J and Hagopian, K. (1962) Some Applications of Two-Dimensional Ionophoresis. Anal Biochern. 3: 276-284
Nguyen DN, Becker GW, Riggin RM (1995) Protein mass spectrometry: applications to analytical
biotechnology. J. Chromatogr. 705: 21-45
Rose K, Savoy LA, Offord RE, Wingfield P (1988) C-terminal peptide identification by fast atom bombardment mass spectrometry. Biochem J. 250: 253-259
Shenoy NR, Shively JE, Bailey JM (1993) Studies in C-terminal Sequencing: New reagents for the synthesis of peptidylthiohydantoins. J. Prot. Chern. 12: 195-205
Stults JT, Lai J, McCune S, Wetzel R (1993) Simplification of high-energy collision spectra of peptides
by amino terminal derivatization. Anal Chern 65: 1703 -1708
Takamoto K, Kamo M, Kubota K, Satake K, Tsugita A (1995) Carboxy-terminal degradation of peptides using perfluoroacyl anhydrides. Eur. J. Biochem 228: 362-372.
Takao T, Gonzalez LJ, Yoshidome K, Sato K, Asada T, Kammei Y, Shimonishi Y. (1993) Automatic
precursor-ion switching in a four sector tandem mass spectrometer and its application to acquisition of the MS/MS product ions derived from a partially 180-labeled peptide for their facile assignments. Anal Chern. 65: 2394-2399
Takao T, Hori H, Okamoto K, Harada A, Kamachi M, Shimonishi Y. (1991) Facile assignment
of sequence ions of a peptide labelled with 18 0 at the carboxyl terminus. Rapid Commun. Mass
Spectrom. 5: 312-315.
Tsugita A, Takamoto K, Kamo M, Iwadate H (1992) C-terminal sequencing of protein. A novel partial
acid hydrolysis and analysis by mass spectrometry. Eur. J. Biochem. 206 : 691- 696
Wetzel R, Halualani R, Stults JT, Quan C (1990) A general method for highly selective cross-linking of
unprotected polypeptides via pH controlled modification of N-terminal a-amino groups. Bioconjugate Chern 1: 114-122
187
Table 12.1. Summary of the experimental masses observed in the isolated C-terminal peptide from
the different proteins characterized in this study
Protein
Cytochrome C
IL-2m
TAB-9
Myoglobin
Streptokinase
Mucorpepsin
References
Molecular
Weight (kDA)
11.7
15.6
16.6
17.2
47.3
57.2
(M+HJ'
theor.
1805.8
1522.9
592.3
1983.8
1631.7
1394.6
(M+Hr
1805.9
1522.8
592.2
1984.1
1633.0
1395.7
Sequence
Mal- 92 EDLIAYLK*K* ATNE I04
Ac- 122 WITFAQSIISTLT I34
AC_ 158 AFVTII6'
SUC_140K*DIAK*YK*ELGYQGI53
Suc-403YTGTPIPDNPNDK*415
Ac- 1221 G FAPLASGYENN 134
Bailey JM, Shively JE (1990) Carboxy-terminal sequencing: Formation and hydrolysis of C-terminal
Peptidylthiohydantoins. Biochemistry 29: 3145 - 3156.
Boyd VL, Bozzini M, Zon G, Noble RL, Mattaliano R (1992) Sequencing of peptides and proteins from
the carboxy terminus. Anal. Biochem. 206: 344-352
Butler PJG, Harris JI, Hartley, Bsleberman R (1969) The use of maleic anhydride for the reversible
blocking of amino groups in polypeptide chains. Biochem J 112: 679-689
Gorman JJ, Shiell BJ (1993) Isolation of carboxyl-termini and blocked amino-termini of viral proteins
by high-performance cation-exchange chromatography J. Chromo 646: 193-205.
Isobe T, Ichimura T, Okuyama T (1986) Identification of the C-Terminal Portion of a Protein by Comparative Peptide Mapping. Anal Biochem. 155: 135-140
Kumazaki T, Nakako T, Arisaka F, Ishii S (1986) A novel method for selective isolation of C-terminal
pep tides from tryptic digests of proteins by immobilized anhydrotrypsin: application to structural
analyses of the tail sheath and tube proteins from bacteriophage T4. Proteins 1: 100 -107.
Miller CG, Bailey JM (1995) Biotechnology applications of C-terminal sequence analysis. Genetic
Engineering News 15: 12-16
Murphy CM, Fenselau C (1995) Recognition of Carboxy-Terminal Peptide in Cyanogen Bromide
Digest of Proteins. Anal Chern. 67: 1644-1647
Naughton, J and Hagopian, K. (1962) Some Applications of Two-Dimensional Ionophoresis. Anal Biochern. 3: 276-284
Nguyen DN, Becker GW, Riggin RM (1995) Protein mass spectrometry: applications to analytical
biotechnology. J. Chromatogr. 705: 21-45
Rose K, Savoy LA, Offord RE, Wingfield P (1988) C-terminal peptide identification by fast atom bombardment mass spectrometry. Biochem J. 250: 253-259
Shenoy NR, Shively JE, Bailey JM (1993) Studies in C-terminal Sequencing: New reagents for the synthesis of peptidylthiohydantoins. J. Prot. Chern. 12: 195-205
Stults JT, Lai J, McCune S, Wetzel R (1993) Simplification of high-energy collision spectra of peptides
by amino terminal derivatization. Anal Chern 65: 1703 -1708
Takamoto K, Kamo M, Kubota K, Satake K, Tsugita A (1995) Carboxy-terminal degradation of peptides using perfluoroacyl anhydrides. Eur. J. Biochem 228: 362-372.
Takao T, Gonzalez LJ, Yoshidome K, Sato K, Asada T, Kammei Y, Shimonishi Y. (1993) Automatic
precursor-ion switching in a four sector tandem mass spectrometer and its application to acquisition of the MS/MS product ions derived from a partially 180-labeled peptide for their facile assignments. Anal Chern. 65: 2394-2399
Takao T, Hori H, Okamoto K, Harada A, Kamachi M, Shimonishi Y. (1991) Facile assignment
of sequence ions of a peptide labelled with 18 0 at the carboxyl terminus. Rapid Commun. Mass
Spectrom. 5: 312-315.
Tsugita A, Takamoto K, Kamo M, Iwadate H (1992) C-terminal sequencing of protein. A novel partial
acid hydrolysis and analysis by mass spectrometry. Eur. J. Biochem. 206 : 691- 696
Wetzel R, Halualani R, Stults JT, Quan C (1990) A general method for highly selective cross-linking of
unprotected polypeptides via pH controlled modification of N-terminal a-amino groups. Bioconjugate Chern 1: 114-122