Proteome of monocytes primed with lipopolysaccharide: analysis of the abundant proteins.
We performed a proteomic analysis of monocytes primed by lipopolysaccharide (LPS) in vitro, using two-dimensional gels stained with Coomassie blue. We found 16 proteins of approximately 500 detected that either increased or decreased in abundance as a result of priming by LPS (14 with P </= 0.05). The proteins were identified by comparing the masses of their tryptic peptides with those of all known proteins, using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and the SWISS-PROT database. Identities were confirmed by matching the sequence of several tryptic peptides, using liquid chromatography electrospray-ionization quadrupole ion trap mass spectrometry. There were increases in the protective enzymes superoxide dismutase and catalase, in four calgranulins, in the cytokine pre-B cell enhancing factor, and in annexin 2, macrophage capping protein, transketolase, pyruvate kinase, and serine/threonine protein kinase 10. Proteins that decreased in abundance were integrin alpha-IIB, protein disulfide isomerase, and platelet-activating factor acetylhydrolase. Many of these altered proteins have interesting functions in inflammation.
Gadgil
Himanshu S
HS
Department of Molecular Sciences, The University of Tennessee Health Science Center, Memphis, USA.
Pabst
Karen M
KM
Giorgianni
Francesco
F
Umstot
Edward S
ES
Desiderio
Dominic M
DM
Beranova-Giorgianni
Sarka
S
Gerling
Ivan C
IC
Pabst
Michael J
MJ
D000595:Amino Acid Sequence; D003720:Densitometry; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D039421:Integrin alpha2 / Q000032:analysis / Q000276:immunology; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009000:Monocytes / Q000737:chemistry* / Q000187:drug effects; D011506:Proteins / Q000032:analysis; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013450:Sulfones / Q000737:chemistry
D039421:Integrin alpha2; D008070:Lipopolysaccharides; D011506:Proteins; D020543:Proteome; D013450:Sulfones; C002010:4-(2-aminoethyl)benzenesulfonylfluoride