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
Dans Zh
| WPRIM
| ID: wpr-316696
Responsable en Bibliothèque :
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.
Texte intégral:
1
Indice:
WPRIM
Sujet Principal:
Sites de fixation
/
Protéines recombinantes
/
Glycoprotéines membranaires
/
Calcium
/
Sélectine P
/
Leucocytes
/
Métabolisme
/
Mutation
Type d'étude:
Prognostic_studies
Limites du sujet:
Humans
langue:
Zh
Texte intégral:
Acta Physiologica Sinica
Année:
2008
Type:
Article