Expression and identification of recombinant P-selectin and P-selectin glycoprotein ligand-1 / 生理学报
Acta Physiologica Sinica
; (6): 520-524, 2008.
Article
in Chinese
| WPRIM (Western Pacific)
| ID: wpr-316696
Responsible library:
WPRO
ABSTRACT
P-selectin, one of the membrane proteins, expresses on platelet and endothelia and interacts with P-selectin glycoprotein ligand-1 (PSGL-1) on leukocyte membrane. This interaction mediates leukocytes rolling on endothelial membrane and then induces leukocyte recruitment to the site of infection or tissue injury. In the present study, we constructed the recombinant wild type human P-selectin, its calcium-binding sites mutants and recombinant PSGL-1-globulin (PSGL-1-Rg). They expressed in Sf9 cells by using the baculovirus expression system and were purified by TalonTM metal or Protein A affinity chromatography. The results showed that the recombinant PSGL-1-Rg interacted with recombinant wild type P-selectin and two P-selectin mutants with 2 calcium-binding sites mutation respectively, but could not bind to the P-selectin mutant with all 4 calcium-binding sites mutation. Therefore, we verified the importance of P-selectin calcium-binding sites for its interaction with PSGL-1.
Full text:
Available
Database:
WPRIM (Western Pacific)
Main subject:
Binding Sites
/
Recombinant Proteins
/
Membrane Glycoproteins
/
Calcium
/
P-Selectin
/
Leukocytes
/
Metabolism
/
Mutation
Type of study:
Prognostic study
Limits:
Humans
Language:
Chinese
Journal:
Acta Physiologica Sinica
Year:
2008
Document type:
Article