Nano-ESI-MS/MS identification on differentiation-associated proteins in M1 mouse myeloid leukemia cells induced by IL-6.
- Author:
Qing XIA
1
;
Hong-xia WANG
;
Jie WANG
;
Bing-yu LIU
;
Mei-ru HU
;
Xue-min ZHANG
;
Bei-fen SHEN
Author Information
- Publication Type:Journal Article
- MeSH: Actin Depolymerizing Factors; Amino Acid Sequence; Animals; Apoptosis; Cell Transformation, Neoplastic; drug effects; Destrin; Interleukin-6; analysis; pharmacology; Leukemia, Myeloid, Acute; metabolism; pathology; Mice; Microfilament Proteins; analysis; Molecular Sequence Data; Nanotechnology; Recombinant Proteins; pharmacology; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; methods; Tumor Suppressor Proteins; analysis
- From: Acta Academiae Medicinae Sinicae 2004;26(5):483-487
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo identify two differentiation-associated proteins induced by rhIL-6 in M1 mouse myeloid leukemia cells.
METHODSProtein spots were excised from 2-D gels and digested in-gel with trypsin. The trypsin lysis products were first analyzed by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) through peptide mass fingerprinting and then performed peptide sequencing by nano-electrospray ionization mass spectrometry/mass spectrometry (nano-ESI-MS/MS). The database search was finished with the Mascot search engine (http://www.matrixscience.co.uk) using the data processed through MaxEnt3 and MasSeq.
RESULTSThe two proteins were not revealed by peptide mass fingerprint using MALDI-TOF-MS, while they were respectively identified as Destrin and Putative protein after the sequence of their trypic peptides were obtained by the nano-ESI-MS/MS techniques.
CONCLUSIONNano-ESI-MS/MS technique can successfully identify the two differentiation-associated proteins induced by rhIL-6 and has great advantage in protein analysis.
