Isolation of a lymphocyte chemotactic factor produced by the murine thymic epithelial cell line MTEC1: identification as a 30 kDa glycosylated form of MCP-1.
A medullary-type murine thymic epithelial cell line (MTEC1) established in our laboratory constitutively produces multiple species of chemotactic factors, which attract lymphocytes, neutrophils, and monocytes. Chemotactic proteins were isolated from MTEC1 supernatant and purified to homogeneity by adsorption to controlled pore glass, heparin-Sepharose chromatography, cation-exchange FPLC and RP-HPLC. A chemotactic factor for both lymphocytes and monocytes was identified as a 30 kDa protein by SDS-PAGE analysis under reducing conditions. After cleavage of the NH2-terminally blocked protein with formic acid, the amino acid sequence of the internal fragment was analysed and found to be identical to the amino acid sequence of mouse MCP-1/JE. The protein is hence identified as a glycosylated natural form of MCP-1/JE secreted by thymic epithelial cells. The 30 kDa glycosylated form of MCP-1 shows lower specific chemotactic activity (CA) for both lymphocytes and monocytes than the 6-7 kDa unglycosylated form of MCP-1.
Liu
Z G
ZG
Department of Immunology, Beijing Medical University, China.
Haelens
A
A
Wuyts
A
A
Struyf
S
S
Pang
X W
XW
Proost
P
P
Chen
W F
WF
van Damme
J
J
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D018932:Chemokine CCL2 / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D004847:Epithelial Cells; D004848:Epithelium / Q000737:chemistry; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006031:Glycosylation; D007375:Interleukin-1 / Q000494:pharmacology; D015850:Interleukin-6 / Q000096:biosynthesis / Q000378:metabolism; D008214:Lymphocytes / Q000187:drug effects; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009000:Monocytes / Q000187:drug effects; D011070:Poly I-C / Q000494:pharmacology; D011499:Protein Processing, Post-Translational; D018108:Respirovirus / Q000502:physiology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013154:Spleen / Q000166:cytology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013950:Thymus Gland / Q000166:cytology*
D018932:Chemokine CCL2; D007375:Interleukin-1; D015850:Interleukin-6; D013755:Tetradecanoylphorbol Acetate; D011070:Poly I-C