Mechanism of LigandInduced Internalization of CCR7

1
Mechanism of Ligand-Induced Internalization of
CCR7

• BMS Cell Biology 260
• May 30, 2002

2
CCR7 is required for adaptive immune responses
Cyster JG. (1999). Science 2862098-102
3
Internalization of CCR7
CCR7
SLC or ELC
ELC
Æ
CCR7
CCR7
SLC or ELC
SLC

4
Endocytosis
Kirchhausen T. (2000). Nature Reviews Molecular
Cell Biology 1 187-198
5
Key Questions

• How does ELC promote selective internalization of
  the receptor?
• How does CCR7 get internalized?

6
Binding of ELC and SLC to CCR7
Measure
mutate

• Binding
• AP-labeled ligands
• Response
• Ca2 flux
• Chemotaxis
• Internalization
• Flow cytometry
• 125I-labeled ligand
• Dimerization

ELC
SLC
NH2
CCR7
COOH
7
Key Questions

• How does ELC promote selective internalization of
  the receptor?
• How does CCR7 get internalized?

8
Clathrin
Kirchhausen T. (2000). Nature Reviews Molecular
Cell Biology 1 187-198
9
Clathrin-mediated endocytosis
Takei et al. (2001). Trends Cell Biol 11 385-91
10
Caveolae EM
Anderson R. (1998). Annu. Rev. Biochem.
67199-225.
11
Adapter Molecules
McDonald et al. (2001). Cell Signal 13683-9
12
Immunofluorescence Microscopy
b2-adrenergic receptor
Dextran
Gagnon et al. (1998). J Biol Chem 273 6976-6981
13
CCR7 is required for adaptive immune responses
Cyster JG. (1999). Science 2862098-102