Sequencing of novel protein from Bacillus pumilus PH-01 using a high-resolution hybrid quadrupole-time-of-flight mass spectrometer
SCIE
SCOPUS
- Title
- Sequencing of novel protein from Bacillus pumilus PH-01 using a high-resolution hybrid quadrupole-time-of-flight mass spectrometer
- Authors
- Chang, YS; Hong, HB; Kim, BH; Hall, SC; Settineri, TA
- Date Issued
- 2001-08-24
- Publisher
- ELSEVIER SCIENCE BV
- Abstract
- Protein identification can be accomplished with an enzymatic digestion of the protein followed by mass spectrometric analysis of the peptide mass fingerprint followed by database searching. However, if a protein is not in a database, sequence information must be obtained to characterize and identify it. This can be done either by classical Edman sequencing or/and by tandem mass spectrometry. To determine the sequence of an unknown protein from Bacillus pumilus PH-01, which adsorbs environmental pollutants such as polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), and biphenyls (PCBs), both sequencing and de novo peptide sequencing by tandem MS/MS of the peptide fragments were performed. Edman sequencing of the reduced and alkylated protein revealed the majority of the sequence; however, all information on disulfide bonding was lost. Therefore, a tryptic digest of the native protein was performed to obtain both complete sequence information and the connectivity of the disulfide bonds. We performed the de novo sequencing using a hybrid quadrupole-time-of-flight mass spectrometer (Q-TOF MS) instrument. The high mass accuracy and sensitivity of the hybrid Q-TOF MS made low-level sequencing of this novel naturally isolated protein possible. (C) 2001 Elsevier Science B.V.
- Keywords
- protein identification; sequencing; protein; Bacillus pumilus; hybrid Q-TOF MS; MS/MS; SOURCE DECAY; PEPTIDE; IDENTIFICATION
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/19405
- DOI
- 10.1016/S1387-3806(01)00463-8
- ISSN
- 1387-3806
- Article Type
- Article
- Citation
- INTERNATIONAL JOURNAL OF MASS SPECTROMETRY, vol. 209, no. 1, page. 47 - 55, 2001-08-24
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