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Published on: May 31, 2018
p161, a murine membrane protein expressed on mast cells and some macrophages, is mouse CD13/aminopeptidase N
1Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
Abstract:
pl6l is a membrane glycoprotein expressed on mast cells and on activated macrophages but on few if any other cells of hematopoietic lineages. Its lack of expression on basophils makes it useful to distinguish mast cells from basophils and aids in the analysis of mast cells and their precursors. p161 was purified from the mast cell line CFTL-12 by affinity chromatography and subjected to limited proteolysis. The sequences of the resultant peptides indicated that p161 is homologous with rat and human CD13/aminopeptidase N. Using oligonucleotide primers derived from rat CD13 cDNA, a mouse cDNA was obtained. Its deduced amino acid sequence displays 87% identity with rat CD13 and 76 % identity with human CD13. Expression of the mouse cDNA in M12 cells, which are p161 negative, renders these cells positive for staining with the monoclonal anti-p161 Ab, K-1. Furthermore, a mAb raised against partially purified mouse intestinal aminopeptidase N specifically blocked the binding of K-1 to both CFTL-12 cells and the transfected M12 cells. These results strongly indicate that mouse p161 is CD13/aminopeptidase N. Northern blot analysis shows that p161 mRNA is most abundantly expressed in the intestinal tract and kidney and is present in liver, lymph node, spleen, and brain.
Insights
Mouse p161, a membrane glycoprotein on mast cells, is identified as CD13/aminopeptidase N. This finding aids in distinguishing mast cells from basophils and analyzing mast cell precursors.
Area of Science:
- Immunology
- Molecular Biology
- Biochemistry
Background:
- p161 is a membrane glycoprotein found on mast cells and activated macrophages.
- Its expression profile is useful for differentiating mast cells from basophils.
Purpose of the Study:
- To identify and characterize the mouse p161 protein.
- To confirm its homology with known CD13/aminopeptidase N.
Main Methods:
- Affinity chromatography and limited proteolysis for p161 purification.
- Peptide sequencing to determine protein homology.
- cDNA cloning and expression in M12 cells.
- Monoclonal antibody (mAb) binding assays.
Main Results:
- p161 peptide sequences showed homology to rat and human CD13/aminopeptidase N.
- Mouse cDNA sequence exhibited high identity (87% rat, 76% human) with CD13.
- Transfection of M12 cells with mouse cDNA resulted in p161 expression.
- A mAb against mouse intestinal aminopeptidase N blocked K-1 binding, confirming p161 identity.
Conclusions:
- Mouse p161 is confirmed to be CD13/aminopeptidase N.
- p161 mRNA is highly expressed in the intestine and kidney, with lower levels in other tissues.

