Abstract
Multiple CC chemokines bind to CCR1, which plays important roles in immune and inflammatory responses. To search for proteins involved in the CCR1 signaling pathway, we screened a yeast two-hybrid library using the cytoplasmic tail of CCR1 as the bait. One of the positive clones contained an open reading frame of 456 bp, of which the nucleotide sequence was identical to that of proteolipid protein 2 (PLP2), also known as protein A4. Mammalian two-hybrid and coimmunoprecipitation analyses demonstrated the association of PLP2/A4 with CCR1. Indirect immunofluorescence analysis revealed that PLP2/A4 was predominantly located in plasma membrane and colocalized with CCR1 in transfected human HEK293 cells. In addition, focal staining of CCR1 appeared on the periphery of the membrane upon short exposure to Leukotactin-1(Lkn-1)/CCL15, a CCR1 agonist, and was costained with PLP2/A4 on the focal regions. PLP2/A4 mRNAs were detected in various cells such as U-937, HL-60, HEK293, and HOS cells. Overexpression of PLP2/A4 stimulated a twofold increase in the agonist-induced migration of HOS/CCR1 cells, implicating a functional role for PLP2/A4 in the chemotactic processes via CCR1.
Original language | English |
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Pages (from-to) | 768-772 |
Number of pages | 5 |
Journal | Biochemical and biophysical research communications |
Volume | 324 |
Issue number | 2 |
DOIs | |
Publication status | Published - 2004 Nov 12 |
Externally published | Yes |
Bibliographical note
Funding Information:This work was supported by SRC fund to IRC at University of Ulsan from the KOSEF and the Korean Ministry of Science and Technology.
Keywords
- CCR1
- Chemokine receptor
- Chemotaxis
- Protein A4
- Protein interaction
- Proteolipid protein 2
ASJC Scopus subject areas
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology