Mass balance strategy for protein sequencing. Application to a protein with an extended DNPNNP repeating motif.
The value of the mass balance strategy is illustrated in the sequence determination of S. aureus V8 protease. Capillary electrophoresis, electrospray mass spectrometry, and high performance tandem mass spectrometry are used as well as proteolysis and Edman degradation. The carboxy terminus is found to contain 17 irregularly repeating units of the triptych motifs NNP and DNP, which provide a challenge to any strategy involving mapping, sequencing, and overlapping of hydrolytic peptides.
Hua
S
S
Department of Chemistry and Biochemistry, University of Maryland Baltimore County.
Vestling
M M
MM
Murphy
C M
CM
Bryant
D K
DK
Height
J J
JJ
Fenselau
C
C
Theibert
J
J
Collins
J H
JH
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D003488:Cyanogen Bromide / Q000737:chemistry; D004140:Dinitrophenols / Q000032:analysis / Q000737:chemistry; D004586:Electrophoresis / Q000379:methods; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009842:Oligopeptides / Q000032:analysis / Q000737:chemistry; D010446:Peptide Fragments / Q000032:analysis; D010449:Peptide Mapping; D011506:Proteins / Q000737:chemistry*; D012091:Repetitive Sequences, Nucleic Acid; D017421:Sequence Analysis / Q000379:methods*; D012697:Serine Endopeptidases / Q000737:chemistry
D004140:Dinitrophenols; D009842:Oligopeptides; D010446:Peptide Fragments; D011506:Proteins; C024118:DNP-peptide; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D003488:Cyanogen Bromide