The use of proteome similarity for the qualitative and quantitative profiling of reperfused myocardium.
An LC-MS-based approach is presented for the identification and quantification of proteins from unsequenced organisms. The method relies on the preservation of homology across species and the similarity in detection characteristics of proteomes in general. Species related proteomes share similarity that progresses from the amino acid frequency distribution to the complete amino sequence of matured proteins. Moreover, the comparative analysis between theoretical and experimental proteome distributions can be used as a measure for the correctness of detection and identification obtained through LC-MS-based schemes. Presented are means to the identification and quantification of rabbit myocardium proteins, immediately after inducing cardiac arrest, using a data-independent LC-MS acquisition strategy. The employed method of acquisition affords accurate mass information on both the precursor and associated product ions, whilst preserving and recording the intensities of the ions. The latter facilitates label-free quantification. The experimental ion density observations obtained for the rabbit sub proteome were found to share great similarity with five other mammalian samples, including human heart, human breast tissue, human plasma, rat liver and a mouse cell line. Redundant, species-homologues peptide identifications from other mammalian organisms were used for initial protein identification, which were complemented with peptide identifications of translated gene sequences. The feasibility and accuracy of label-free quantification of the identified peptides and proteins utilizing above mentioned strategy is demonstrated for selected cardiac rabbit proteins.
Vissers
Johannes P C
JP
Waters Corporation, Atlas Park, Simonsway, Manchester M22 5PP, United Kingdom. hans_vissers@waters.com
Pons
Sandrine
S
Hulin
Anne
A
Tissier
Renaud
R
Berdeaux
Alain
A
Connolly
Joanne B
JB
Langridge
James I
JI
Geromanos
Scott J
SJ
Ghaleh
Bijan
B
D000595:Amino Acid Sequence; D000818:Animals; D002853:Chromatography, Liquid; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D015425:Myocardial Reperfusion; D009206:Myocardium / Q000737:chemistry*; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D020543:Proteome; D011817:Rabbits; D015203:Reproducibility of Results
D011506:Proteins; D020543:Proteome