Table 1.
Amino acid sequence of peptides obtained by digestion with trypsin, chymotrypsin or Glu-C endoproteinases from Peak 1 and Peak 2 of RP-HPLC, as determined by mass spectrometry (MALDI-TOF-TOF or nESI-MS/MS) or Edman N-terminal sequencing.
| Peptide Sequence | HPLC Peak | m/z | z | Confidence a | Error b (ppm) | Method c | Domain |
|---|---|---|---|---|---|---|---|
| YIELVIVADHR | 1 | 1327.8 | 1 | MV | 47.4 | MALDI/DE NOVO | metalloproteinase |
| NLTPEEQRY | 1 | 1149.6 | 1 | MV | 40.0 | MALDI/DE NOVO | metalloproteinase |
| SRYHFVANR | 1 | 1149.6 | 1 | MV | 7.8 | MALDI/DE NOVO | metalloproteinase |
| YHFVANR | 1 | - | MV | - | EDMAN | metalloproteinase | |
| MAHELGHNLGIHQDR | 1 | - | - | MV | - | EDMAN | metalloproteinase |
| MAHELGHNLGIHQDR | 1 | 1727.8 | 1 | MV | 22.5 | MALDI/DE NOVO | metalloproteinase |
| MAHELGHNLGLHQDR | 1 | 576.6180 | 3 | 99 | 0.2 | nESI/DE NOVO | metalloproteinase |
| DSCSCGSNSCIMSATVSNEPSSR | 1 | 2493.0 | 1 | MV | 10.0 | MALDI/DE NOVO | metalloproteinase |
| CIDNEPLR | 1 | 1016.5 | 1 | MV | 16.7 | MALDI/DE NOVO | metalloproteinase |
| NEPLRTDIVSPPFCGNYYPE * | 1 | 2368.3 | 1 | MV | 88.2 | MALDI/DE NOVO | metalloproteinase/disintegrin |
| LTTGSQCAEGLCCDQCR | 1 | 2015.9 | 1 | MV | 47.6 | MALDI/DE NOVO | disintegrin |
| FEGLCCDQCR | 1 | 1344.4 | 1 | MV | 84.0 | MALDI/DE NOVO | disintegrin |
| RTDIVSPPF ** | 1 | 1031.6 | 1 | MV | 46.5 | MALDI/DE NOVO | disintegrin |
| YIELVIVADHR | 2 | 1327.8 | 1 | MV | 47.4 | MALDI/DE NOVO | metalloproteinase |
| VTLSADDTLDLFGTWR | 2 | - | MV | - | EDMAN | metalloproteinase | |
| VALIGLEIWSSGELSK | 2 | 1702.0 | 1 | MV | 34.0 | MALDI/DE NOVO | metalloproteinase |
| YHFVANR | 2 | 906.5 | 1 | MV | 46.3 | MALDI/DE NOVO | metalloproteinase |
| MAHDLGHNLCIDHDR | 2 | 1803.9 | 1 | MV | 54.8 | MALDI/DE NOVO | metalloproteinase |
| MAHDLGHNLCLDHDR | 2 | 902.4039 | 2 | 99 | −0.4 | nESI/DE NOVO | metalloproteinase |
| MAHDLGHNLCLDHDDR | 2 | 959.9163 | 2 | 99 | −1.5 | nESI/DE NOVO | metalloproteinase |
| MAHELGHNLGLHQDDR | 2 | 614.9611 | 3 | 99 | 1.2 | nESI/DE NOVO | metalloproteinase |
| MAHELGHNLGLHQDR | 2 | 864.4214 | 2 | 99 | −2.1 | nESI/DE NOVO | metalloproteinase |
| NKPSETDIVSPPVCGNYY ** | 2 | 2040.1 | 1 | MV | 79.4 | MALDI/DE NOVO | metalloproteinase/disintegrin |
| CNGISAGCPRNPF ** | 2 | 1449.7 | 1 | MV | 44.1 | MALDI/DE NOVO | disintegrin |
| RARGDDVNDYCNGISAGCPRNPFH * | 2 | 2748.4 | 1 | MV | 68.7 | MALDI/DE NOVO | disintegrin |
| ARGDDVNDYCNGISAGCPR | 2 | 2097.1 | 1 | MV | 101.5 | MALDI/DE NOVO | disintegrin |
| ARGDDVDDYCDGLDAGCPR | 2 | 709.6204 | 3 | 91 | −1.7 | nESI/DE NOVO | disintegrin |
| AATCKLTTGSQCADGLCCDQCKFMRE * | 2 | 3068.5 | 1 | MV | 71.0 | MALDI/DE NOVO | disintegrin |
a Confidence: MALDI/DE NOVO and EDMAN (MV, manual validation); nESI/DE NOVO (PEAKS 8 De Novo average local confidence); b Error: error of the precursor ion (in ppm) relative to the proposed sequence; c Method: MALDI/DE NOVO: MALDI-TOF-TOF and manual de novo sequencing; EDMAN: N-terminal sequencing; nESI/DE NOVO: nLC-nanoelectrospray MS/MS and automatic de novo sequencing using PEAKS 8 software; * peptides generated by Glu-C endoproteinase digestion; ** peptides generated by chymotrypsin digestion. Peptides without asterisks were obtained following trypsin digestion. All cysteines are carboxamidomethylated. Sequences colored in red correspond to the mutated catalytic site.