Determinants in the cytoplasmic domain of P-selectin required for sorting to secretory granules - PubMed (original) (raw)

Determinants in the cytoplasmic domain of P-selectin required for sorting to secretory granules

P W Modderman et al. Biochem J. 1998.

Abstract

P-selectin is a granule membrane protein of platelets and endothelial cells that is expressed at the plasma membrane after cell activation. To determine which residues in its cytoplasmic tail are important for sorting to storage granules during biosynthesis, we expressed P-selectin mutants in AtT-20, a murine cell line with secretory granules that contain the hormone corticotropin ('ACTH'). Immunofluorescence microscopy of permeabilized cells revealed that wild-type P-selectin and mutants with alanine substitutions at 14 different positions in the cytoplasmic tail were concentrated in the tips of the cellular processes, which contain the majority of corticotropin granules. However, targeting to the cell tips was greatly decreased for Tyr777-->Ala, Tyr777-->Phe, Gly778-->Ala, Phe780-->Ala and Leu768/Asn769-->Ala/Ala mutants. The reduced presence of these mutants in corticotropin granules was confirmed by immunoelectron microscopy. Stimulation of AtT-20 transfectants with 8-Br-cAMP resulted in a significant increase in membrane expression of wild-type P-selectin, but in only a marginal increase in the surface expression of the five mutants. Antibody binding studies with intact and permeabilized cells demonstrated that the percentage of P-selectin that is expressed on the surface of the cells was considerably higher for these mutants than for wild-type P-selectin (6%), ranging from approximately 20% for the Gly778 and Phe780 mutants to 63% for the Leu768/Asn769 mutant. Taken together, these results indicate that Tyr777, Gly778 and Phe780 form part of an atypical tyrosine-based motif, which also requires the presence Leu768 and/or Asn769 to mediate sorting of P-selectin to secretory granules.

PubMed Disclaimer

Similar articles

• P-selectin, a granule membrane protein of platelets and endothelial cells, follows the regulated secretory pathway in AtT-20 cells.
  Koedam JA, Cramer EM, Briend E, Furie B, Furie BC, Wagner DD. Koedam JA, et al. J Cell Biol. 1992 Feb;116(3):617-25. doi: 10.1083/jcb.116.3.617. J Cell Biol. 1992. PMID: 1370497 Free PMC article.
• Cytoplasmic domain of P-selectin (CD62) contains the signal for sorting into the regulated secretory pathway.
  Disdier M, Morrissey JH, Fugate RD, Bainton DF, McEver RP. Disdier M, et al. Mol Biol Cell. 1992 Mar;3(3):309-21. doi: 10.1091/mbc.3.3.309. Mol Biol Cell. 1992. PMID: 1378326 Free PMC article.
• AP-3 adaptor functions in targeting P-selectin to secretory granules in endothelial cells.
  Daugherty BL, Straley KS, Sanders JM, Phillips JW, Disdier M, McEver RP, Green SA. Daugherty BL, et al. Traffic. 2001 Jun;2(6):406-13. doi: 10.1034/j.1600-0854.2001.002006406.x. Traffic. 2001. PMID: 11389768
• The cytoplasmic domain of P-selectin contains a sorting determinant that mediates rapid degradation in lysosomes.
  Green SA, Setiadi H, McEver RP, Kelly RB. Green SA, et al. J Cell Biol. 1994 Feb;124(4):435-48. doi: 10.1083/jcb.124.4.435. J Cell Biol. 1994. PMID: 7508941 Free PMC article.
• Residues throughout the cytoplasmic domain affect the internalization efficiency of P-selectin.
  Setiadi H, Disdier M, Green SA, Canfield WM, McEver RP. Setiadi H, et al. J Biol Chem. 1995 Nov 10;270(45):26818-26. doi: 10.1074/jbc.270.45.26818. J Biol Chem. 1995. PMID: 7592923

Cited by

• Storage of factor VIII variants with impaired von Willebrand factor binding in Weibel-Palade bodies in endothelial cells.
  van den Biggelaar M, Bouwens EA, Voorberg J, Mertens K. van den Biggelaar M, et al. PLoS One. 2011;6(8):e24163. doi: 10.1371/journal.pone.0024163. Epub 2011 Aug 31. PLoS One. 2011. PMID: 21909383 Free PMC article.
• Peritoneal macrophages express both P-selectin and PSGL-1.
  Tchernychev B, Furie B, Furie BC. Tchernychev B, et al. J Cell Biol. 2003 Dec 8;163(5):1145-55. doi: 10.1083/jcb.200310079. J Cell Biol. 2003. PMID: 14662752 Free PMC article.
• Selective and signal-dependent recruitment of membrane proteins to secretory granules formed by heterologously expressed von Willebrand factor.
  Blagoveshchenskaya AD, Hannah MJ, Allen S, Cutler DF. Blagoveshchenskaya AD, et al. Mol Biol Cell. 2002 May;13(5):1582-93. doi: 10.1091/mbc.01-09-0462. Mol Biol Cell. 2002. PMID: 12006654 Free PMC article.
• Rapid transport of internalized P-selectin to late endosomes and the TGN: roles in regulating cell surface expression and recycling to secretory granules.
  Straley KS, Green SA. Straley KS, et al. J Cell Biol. 2000 Oct 2;151(1):107-16. doi: 10.1083/jcb.151.1.107. J Cell Biol. 2000. PMID: 11018057 Free PMC article.
• Two conserved amino acid motifs mediate protein targeting to the micronemes of the apicomplexan parasite Toxoplasma gondii.
  Di Cristina M, Spaccapelo R, Soldati D, Bistoni F, Crisanti A. Di Cristina M, et al. Mol Cell Biol. 2000 Oct;20(19):7332-41. doi: 10.1128/MCB.20.19.7332-7341.2000. Mol Cell Biol. 2000. PMID: 10982850 Free PMC article.

References

1. 1. Blood. 1996 Feb 15;87(4):1432-8 - PubMed
2. 1. J Exp Med. 1996 Jul 1;184(1):81-92 - PubMed
3. 1. Annu Rev Cell Biol. 1993;9:129-61 - PubMed
4. 1. Mol Biol Cell. 1992 Mar;3(3):309-21 - PubMed
5. 1. Cell. 1995 Jul 28;82(2):279-86 - PubMed

Publication types

MeSH terms

Substances

LinkOut - more resources

• Full Text Sources

  • Europe PubMed Central
  • PubMed Central
  • Silverchair Information Systems