title: Peptide mapping of recombinant human parathyroid hormone by enzymatic digestion and subsequent fast-atom bombardment mass spectrometry.
abstract: Peptide maps of recombinant human parathyroid hormone (rhPTH) were determined by both trypsin and V-8 protease digestion with subsequent fast-atom bombardment mass spectrometry (FAB-MS). Coverage of the sequence was 85% when using trypsin and 90% when using V-8 protease. Five rhPTH variants that were recombinantly produced as models of Asn deamidated type degradation products were measured, and molecular weight differences between their respective deamidated peptide fragments were completely detected. In the V-8 protease digests of some variants, characteristic peptide ions caused by the deamidation were observed and this greatly facilitated the assignment and recognition of the deamidated position. Our data suggest that FAB-mapping of rhPTH via the protease digestion methods used, appears to have great potential for structural investigations of the peptide.
authors.affiliation: Laboratory for Preliminary Drug Evaluation, Chugai Pharmaceutial Co., Ltd, Shizuoka, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Nabuchi
authors.forename: H
authors.initials: H
authors.lastname: Kuboniwa
authors.forename: H
authors.initials: H
authors.lastname: Takasu
authors.forename: Y
authors.initials: Y
authors.lastname: Asoh
authors.forename: H
authors.initials: H
authors.lastname: Ushio
mesh_terms: D000595:Amino Acid Sequence; D006801:Humans; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010281:Parathyroid Hormone / Q000737:chemistry*; D010449:Peptide Mapping / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry; D012697:Serine Endopeptidases; D016339:Spectrometry, Mass, Fast Atom Bombardment; D014357:Trypsin
chemical_list: D010281:Parathyroid Hormone; D011994:Recombinant Proteins; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D014357:Trypsin