| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Regular Article |
a Department of Obstetrics and Gynecology, Department of Cell Biology, Duke University Medical Center, Durham, North Carolina 27710
Formation of the egg's extracellular matrix, the zona pellucida, is critical for fertilization and development of growing embryos. Zona pellucida glycoproteins, ZP1, ZP2, and ZP3, are secreted to form an insoluble extracellular matrix surrounding mammalian eggs. All cloned mammalian zona pellucida sequences contain a furin consensus cleavage site, RXK/RR, upstream of a putative transmembrane domain, which suggests processing by an endoprotease of the furin-proprotein-convertase family. Recombinant expression of human (h) ZP1, ZP2, and ZP3 produces glycoproteins that are secreted and have migration patterns in SDS-PAGE identical to those of native human zona pellucida proteins. Because a C-terminal epitope tag that is present in the cell-associated zona proteins is, however, absent from the secreted zona proteins, secreted recombinant zona pellucida proteins lack their C-terminal regions. Three different strategies were used to explore processing events in the C-terminal region: site-directed mutagenesis of the furin cleavage site, treatment with a competitive inhibitor of all furin family members, and interference with Golgi modifications by Brefeldin A. All treatments altered the SDS-PAGE migration of recombinant hZP3, concordant with cleavage by a furin family member and Golgi glycosylation of secreted hZP3. Furthermore, cleavage of cell-associated hZP3 by exogenous furin converts the migration of cell-associated hZP3 to that of secreted hZP3. To determine whether a similar cleavage pattern exists in zona pellucida proteins that are assembled in the zona matrix, "hZP3 rescue" mouse zonae pellucidae were employed. Immunoblotting experiments revealed that hZP3, assembled and functional in the "hZP3 rescue" mouse zona pellucida, lacks the furin cleavage site, supporting the hypothesis that formation of the zona pellucida matrix involves regulated proteolysis by a member of the furin convertase family.
1 Supported by NIH RO1 HD18201.
2 Correspondence: Patricia Saling, 234 Sands Building, Research Drive, Duke University Medical Center, Durham, NC 27710. FAX: 919 681 6494; patricia.saling{at}duke.edu
3 Current address: Washington University School of Medicine, Box 8126, Renal Division, 660 S. Euclid Avenue, St. Louis, MO 63110.
This article has been cited by other articles:
|
|
 |
|
  J. E. Swain and T. B. Pool ART failure: oocyte contributions to unsuccessful fertilization Hum. Reprod. Update, September 1, 2008; 14(5): 431 - 446. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. C. Darie, M. L. Biniossek, M. A. Gawinowicz, Y. Milgrom, J. O. Thumfart, L. Jovine, E. S. Litscher, and P. M. Wassarman Mass Spectrometric Evidence That Proteolytic Processing of Rainbow Trout Egg Vitelline Envelope Proteins Takes Place on the Egg J. Biol. Chem., November 11, 2005; 280(45): 37585 - 37598. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Yonezawa, K. Kudo, H. Terauchi, S. Kanai, N. Yoda, M. Tanokura, K. Ito, K.-i. Miura, T. Katsumata, and M. Nakano Recombinant Porcine Zona Pellucida Glycoproteins Expressed in Sf9 Cells Bind to Bovine Sperm but Not to Porcine Sperm J. Biol. Chem., May 27, 2005; 280(21): 20189 - 20196. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Quesada, L. M. Sanchez, J. Alvarez, and C. Lopez-Otin Identification and Characterization of Human and Mouse Ovastacin: A NOVEL METALLOPROTEINASE SIMILAR TO HATCHING ENZYMES FROM ARTHROPODS, BIRDS, AMPHIBIANS, AND FISH J. Biol. Chem., June 18, 2004; 279(25): 26627 - 26634. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Jovine, H. Qi, Z. Williams, E. S. Litscher, and P. M. Wassarman A duplicated motif controls assembly of zona pellucida domain proteins PNAS, April 20, 2004; 101(16): 5922 - 5927. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Hoodbhoy and J. Dean Insights into the molecular basis of sperm-egg recognition in mammals Reproduction, April 1, 2004; 127(4): 417 - 422. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Zhao, L. Gold, H. Dorward, L.-f. Liang, T. Hoodbhoy, E. Boja, H. M. Fales, and J. Dean Mutation of a Conserved Hydrophobic Patch Prevents Incorporation of ZP3 into the Zona Pellucida Surrounding Mouse Eggs Mol. Cell. Biol., December 15, 2003; 23(24): 8982 - 8991. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Sasanami, A. M. Hanafy, M. Toriyama, and M. Mori Variant of Perivitelline Membrane Glycoprotein ZPC of Japanese Quail (Coturnix japonica) Lacking Its Cytoplasmic Tail Exhibits the Retention in the Endoplasmic Reticulum of Chinese Hamster Ovary (CHO-K1) Cells Biol Reprod, October 1, 2003; 69(4): 1401 - 1407. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. S. Boja, T. Hoodbhoy, H. M. Fales, and J. Dean Structural Characterization of Native Mouse Zona Pellucida Proteins Using Mass Spectrometry J. Biol. Chem., September 5, 2003; 278(36): 34189 - 34202. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Sasanami, M. Toriyama, and M. Mori Carboxy-Terminal Proteolytic Processing at a Consensus Furin Cleavage Site Is a Prerequisite Event for Quail ZPC Secretion Biol Reprod, May 1, 2003; 68(5): 1613 - 1619. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
