Baugher Paige J, Richmond Ann
Department of Veterans Affairs, Veterans Affairs Medical Center, Nashville, Tennessee 37212-2637, USA.
J Biol Chem. 2008 Nov 7;283(45):30868-78. doi: 10.1074/jbc.M804054200. Epub 2008 Aug 27.
Adaptor protein interaction with specific peptide motifs found within the intracellular, carboxyl terminus of chemokine receptor CXCR2 has been shown to modulate intracellular trafficking and receptor function. Efficient ligand-induced internalization of this receptor is dependent on the binding of adaptor protein 2 to the specific LLKIL motif found within the carboxyl terminus (1). In this study we show that the carboxyl-terminal type 1 PDZ ligand motif (-STTL) of CXCR2 plays an essential role in both proper intracellular receptor trafficking and efficient cellular chemotaxis. First, we show that CXCR2 is sorted to and degraded in the lysosome upon long-term ligand stimulation. We also show that receptor degradation is not dependent upon receptor ubiquitination, but is instead modulated by the carboxyl-terminal type I PDZ ligand of CXCR2. Deletion of this ligand results in increased degradation, earlier co-localization with the lysosome, and enhanced sorting to the Rab7-positive late endosome. We also show that deletion of this ligand effects neither receptor internalization nor receptor recycling. Furthermore, we demonstrate that deletion of the PDZ ligand motif results in impaired chemotactic response. The data presented here demonstrate that the type I PDZ ligand of CXCR2 acts to both delay lysosomal sorting and facilitate proper chemotactic response.
衔接蛋白与趋化因子受体CXCR2细胞内羧基末端特定肽基序的相互作用已被证明可调节细胞内运输和受体功能。该受体高效的配体诱导内化依赖于衔接蛋白2与羧基末端特定的LLKIL基序的结合(1)。在本研究中,我们表明CXCR2的羧基末端1型PDZ配体基序(-STTL)在细胞内受体的正常运输和有效的细胞趋化作用中均发挥着重要作用。首先,我们表明长期配体刺激后,CXCR2会被分选至溶酶体并在其中降解。我们还表明受体降解不依赖于受体泛素化,而是由CXCR2的羧基末端I型PDZ配体调节。该配体的缺失会导致降解增加、与溶酶体更早地共定位以及向Rab7阳性晚期内体的分选增强。我们还表明该配体的缺失既不影响受体内化也不影响受体再循环。此外,我们证明PDZ配体基序的缺失会导致趋化反应受损。此处呈现的数据表明CXCR2的I型PDZ配体既能延迟溶酶体分选又能促进正常的趋化反应。