title: Investigation of bn-44 peptide fragments using high resolution mass spectrometry and isotope labeling.
abstract: An N-terminal deuterohemin-containing hexapeptide (DhHP-6) was designed as a short peptide cytochrome c (Cyt c) mimetic to study the effect of N-terminal charge on peptide fragmentation pathways. This peptide gave different dissociation patterns than normal tryptic peptides. Upon collision-induced dissociation (CID) with an ion trap mass spectrometer, the singly charged peptide ion containing no added proton generated abundant and characteristic bn-44 ions instead of bn-28 (an) ions. Studies by high resolution mass spectrometry (HRMS) and isotope labeling indicate that elimination of 44 Da fragments from b ions occurs via two different pathways: (1) loss of CH3CHO (44.0262) from a Thr side chain; (2) loss of CO2 (43.9898) from the oxazolone structure in the C-terminus. A series of analogues were designed and analyzed. The experimental results combined with Density Functional Theory (DFT) calculations on the proton affinity of the deuteroporphyrin demonstrate that the production of these novel bn-44 ions is related to the N-terminal charge via a charge-remote rather than radical-directed fragmentation pathway.
authors.affiliation: College of Chemistry, Jilin University, Changchun, 130012, China.
authors.forename: Bing
authors.initials: B
authors.lastname: Wang
authors.forename: Jiayi
authors.initials: J
authors.lastname: Yu
authors.forename: Huixin
authors.initials: H
authors.lastname: Wang
authors.forename: Zhonglin
authors.initials: Z
authors.lastname: Wei
authors.forename: Xinhua
authors.initials: X
authors.lastname: Guo
authors.forename: Zhaohui
authors.initials: Z
authors.lastname: Xiao
authors.forename: Zhoufang
authors.initials: Z
authors.lastname: Zeng
authors.forename: Wei
authors.initials: W
authors.lastname: Kong
mesh_terms: D045304:Cytochromes c / Q000737:chemistry; D006427:Hemin / Q000031:analogs & derivatives* / Q000737:chemistry; D007477:Ions / Q000737:chemistry; D007553:Isotope Labeling / Q000379:methods*; D009842:Oligopeptides / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D007477:Ions; D009842:Oligopeptides; D010446:Peptide Fragments; C552681:deuterohemin-alanyl-histidyl-threonyl-valyl-glutamyl-lysine; D006427:Hemin; D045304:Cytochromes c