[Nano-ESI-MS/MS identification on differentiation-associated proteins in M1 mouse myeloid leukemia cells induced by IL-6].
OBJECTIVE
To identify two differentiation-associated proteins induced by rhIL-6 in M1 mouse myeloid leukemia cells.


METHODS
Protein 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.


RESULTS
The 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.


CONCLUSIONS
Nano-ESI-MS/MS technique can successfully identify the two differentiation-associated proteins induced by rhIL-6 and has great advantage in protein analysis.
Xia
Qing
Q
Department of Molecular Immunology, Institute of Basic Medical Science, Academy of Military Medical Sciences, Beijing 100850, China.
Wang
Hong-xia
HX
Wang
Jie
J
Liu
Bing-yu
BY
Hu
Mei-ru
MR
Zhang
Xue-min
XM
Shen
Bei-fen
BF
D051339:Actin Depolymerizing Factors; D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis; D002471:Cell Transformation, Neoplastic / Q000187:drug effects; D051345:Destrin; D015850:Interleukin-6 / Q000032:analysis* / Q000494:pharmacology; D015470:Leukemia, Myeloid, Acute / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D008840:Microfilament Proteins / Q000032:analysis; D008969:Molecular Sequence Data; D036103:Nanotechnology; D011994:Recombinant Proteins / Q000494:pharmacology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D025521:Tumor Suppressor Proteins / Q000032:analysis
D051339:Actin Depolymerizing Factors; D051345:Destrin; C495825:Dstn protein, mouse; D015850:Interleukin-6; D008840:Microfilament Proteins; D011994:Recombinant Proteins; D025521:Tumor Suppressor Proteins