title: Direct N-terminal sequencing of polypeptides using a thermostable bacterial aminopeptidase and MALDI-TOF mass spectrometry.
abstract: Mass spectrometry-based amino acid sequencing is currently based almost entirely on collision-induced peptide fragmentation and analyses. Here, we describe a single-stage MS-based technique for amino acid sequencing involving partial, heterogenous digestion of a peptide by a processive, non-specific, heat-loving Bacillus subtilis-derived aminopeptidase (BsuAP), which acts optimally at 70 °C and allows 'single-shot' sequencing to be carried out through simultaneous accumulation, and detection of sub-populations of peptides of progressively reducing length.
authors.affiliation: Center for Protein Science, Design and Engineering (CPSDE), Department of Biological Sciences, Indian Institute of Science Education & Research (IISER) Mohali, Knowledge City, Sector-81, SAS Nagar, Punjab, 140306, India.
authors.forename: Nitin
authors.initials: N
authors.lastname: Kishor
authors.affiliation: Center for Protein Science, Design and Engineering (CPSDE), Department of Biological Sciences, Indian Institute of Science Education & Research (IISER) Mohali, Knowledge City, Sector-81, SAS Nagar, Punjab, 140306, India. Electronic address: guptasarma@iisermohali.ac.in.
authors.forename: Purnananda
authors.initials: P
authors.lastname: Guptasarma
mesh_terms: D000626:Aminopeptidases / Q000235:genetics / Q000378:metabolism*; D001412:Bacillus subtilis / Q000201:enzymology*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004795:Enzyme Stability; D006358:Hot Temperature; D007194:India; D008970:Molecular Weight; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism*; D059748:Proteolysis; D011994:Recombinant Proteins; D020539:Sequence Analysis, Protein / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D001426:Bacterial Proteins; D009842:Oligopeptides; D010446:Peptide Fragments; D011994:Recombinant Proteins; D000626:Aminopeptidases
keywords: Bacillus subtilis aminopeptidase; MALDI-TOF mass spectrometry; Non-specific aminopeptidase; Peptide N-terminal sequencing; Processive aminopeptidase; Thermostable aminopeptidase