Exploring the platelet proteome via combinatorial, hexapeptide ligand libraries.
A combinatorial ligand library, composed of millions of diverse hexapeptide baits, able to capture and concentrate the "low-abundance" proteome while drastically cutting the concentration of the most abundant species, has been applied to the exploration of the soluble platelet proteome. Mass spectrometry analysis of untreated and library-treated platelets has resulted in the identification of 435 unique gene products. Of those, 147 entries (35% of the total) have not been described among the list of >1100 proteins in proteomic platelet investigations reported before. In addition, the analysis of excised spots from two-dimensional electrophoresis analysis allowed 57 other proteins to be added that were not found in LC-MS analysis, 33 of them not described before in proteomics studies, bringing the total number of new gene products to 180. Thus, the present data add a non-negligible number of species for continuing the "cartography" of the proteomic asset of platelets, in view of completing the mapping procedure for a deeper understanding of the physiology and pathology of this blood cell. Because the capturing process is performed under physiological conditions, by exploiting, for binding to the combinatorial library, the native protein configuration, the described technique is not adapted to capture highly hydrophobic proteins, which need strong denaturing and solubilizing conditions that are incompatible with our working procedure. Thus, our list reports essentially hydrophilic proteins, with negative GRAVY indexes.
Guerrier
Luc
L
Bio-Rad Laboratories, c/o CEA-Saclay, 91191 Gif-sur-Yvette Cedex, France.
Claverol
Stephane
S
Fortis
Frederic
F
Rinalducci
Sara
S
Timperio
Anna Maria
AM
Antonioli
Paolo
P
Jandrot-Perrus
Martine
M
Boschetti
Egisto
E
Righetti
Pier Giorgio
PG
D001792:Blood Platelets / Q000378:metabolism*; D001798:Blood Proteins / Q000737:chemistry*; D020650:Combinatorial Chemistry Techniques / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D005786:Gene Expression Regulation; D006801:Humans; D008024:Ligands; D019151:Peptide Library; D010449:Peptide Mapping; D010455:Peptides / Q000737:chemistry; D040081:Protein Array Analysis; D011485:Protein Binding; D040901:Proteomics / Q000379:methods*
D001798:Blood Proteins; D008024:Ligands; D019151:Peptide Library; D010455:Peptides