Characterization of purified c-type heme-containing peptides and identification of c-type heme-attachment sites in Shewanella oneidenis cytochromes using mass spectrometry.
We describe methods for mass spectrometric identification of heme-containing peptides from c-type cytochromes that contain the CXXCH (X=any amino acid) sequence motif. The heme fragment ion yielded the most abundant MS/MS peak for standard heme-containing peptides with one amino acid difference for both 2+ and 3+ peptide charge states; both sequence and charge affect the extent of heme loss. Application to Shewanella oneidenis demonstrated the utility of this approach for identifying c-type heme-containing peptides from complex proteome samples.
Yang
Feng
F
Biological Sciences Division and Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, P.O. Box 999, Richland, Washington 99352, USA.
Bogdanov
Bogdan
B
Strittmatter
Eric F
EF
Vilkov
Andrey N
AN
Gritsenko
Marina
M
Shi
Liang
L
Elias
Dwayne A
DA
Ni
Shuisong
S
Romine
Margaret
M
Pasa-Tolić
Ljiljana
L
Lipton
Mary S
MS
Smith
Richard D
RD
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000032:analysis*; D001665:Binding Sites; D003574:Cytochrome c Group / Q000032:analysis*; D006418:Heme; D013058:Mass Spectrometry; D010446:Peptide Fragments / Q000302:isolation & purification; D020592:Shewanella / Q000737:chemistry*
D001426:Bacterial Proteins; D003574:Cytochrome c Group; D010446:Peptide Fragments; D006418:Heme