The sea urchin egg yolk granule is a storage compartment for HCL-32, an extracellular matrix protein

1998 ◽  
Vol 76 (1) ◽  
pp. 83-88 ◽  
Author(s):  
Janice Mayne ◽  
John J Robinson
Keyword(s):  
Extracellular Matrix ◽  
Sea Urchin ◽  
Blot Analysis ◽  
Extracellular Matrix Protein ◽  
Yolk Granule ◽  
Cortical Granules ◽  
Yolk Granules ◽  
Storage Compartment ◽  
Protein Gel ◽  
Stage Of Development

We have utilized protein gel blot analysis and immunogold labelling to define the intracellular storage compartment for HCL-32, a 32-kDa protein component of the sea urchin embryonic extracellular matrices, the hyaline layer and basal lamina. Anti-HCL-32 antiserum specifically labelled yolk granules in unfertilized eggs. Cortical granules, mitochondria, sparse granules, and lipid vacuoles were not labelled. Label continued to be detected in the yolk granules through to the blastula stage of development. However, by the gastrula stage no labelling was detected in the yolk granules. In protein gel blot analysis HCL-32 was detected in yolk granules prepared from unfertilized eggs. These results clearly define the yolk granule as a storage compartment for HCL-32, an extracellular matrix protein.Key words: embryo, yolk granule, extracellular matrix.

The hyaline layer in early development of the sea urchin: Cell-extracellular matrix interactions

1989 ◽  
Vol 47 ◽  
pp. 836-837
Author(s):  
David R. McClay ◽  
Mark C. Alliegro ◽  
Steven D Black
Keyword(s):  
Extracellular Matrix ◽  
Early Development ◽  
Sea Urchin ◽  
Protective Layer ◽  
Distinct Group ◽  
Cortical Granules ◽  
Matrix Interactions ◽  
Apical Vesicles ◽  
Double Label ◽  
Extracellular Matrix Interactions

The hyaline layer is released by the sea urchin zygote starting about 30 seconds after fertilization. This layer is elevated above the surface of the zygote where it serves as a protective layer, a barrier to polyspermy, and as an adhesive substrate for the early stages of development. We have been interested in learning how the hyaline layer is stored in the egg, how it is released after fertilization, and how it functions as an extracellular matrix during early development. The layer contains about fifteen abundant proteins plus a number of others that may be present or may contaminate preparations of the layer during isolation procedures. Our goal was to identify the major components of the hyaline membrane, to follow assembly of the structure after fertilization, and to identify molecules that are active as adhesive substrates for cells.Using double-label immunofluorescence and ultrastructural immunogold localization of polyclonal and monoclonal antibodies, the components of the hyaline layer have been localized to four vesicle classes in the unfertilized egg. Centrifugation studies on unfertilized eggs and double-label immunofluorescent studies have shown that the four classes each contain a distinct group of proteins. The four vesicles were followed through oogenesis to learn when proteins of each class were synthesized by the oocyte. Each of the four classes (cortical granules, basal lamina vesicles, apical vesicles and echinonectin vesicles) were synthesized, localized, and relocalized in the oocyte by unique spatial and temporal pathways.

A new extracellular matrix protein of the sea urchin embryo with properties of a substrate adhesion molecule

1992 ◽  
Vol 201 (3) ◽  
pp. 173-178 ◽  
Author(s):  
Valeria Matranga ◽  
Daniela Ferrol ◽  
Francesca Zito ◽  
Melchiorre Cervello ◽  
Eizo Nakano
Keyword(s):  
Extracellular Matrix ◽  
Sea Urchin ◽  
Adhesion Molecule ◽  
Matrix Protein ◽  
Extracellular Matrix Protein ◽  
Substrate Adhesion ◽  
Sea Urchin Embryo

The extracellular matrix of echinoderm and amphibian eggs: Visualization in quick-frozen, deep-etched, and rotary-shadowed specimens

1990 ◽  
Vol 48 (3) ◽  
pp. 60-61
Author(s):  
Barry Bonnell ◽  
Carolyn Larabell ◽  
Douglas Chandler
Keyword(s):  
Extracellular Matrix ◽  
Plasma Membrane ◽  
Sea Urchin ◽  
Crystalline Structure ◽  
Cortical Granule ◽  
Two Dimensional ◽  
Small Peptides ◽  
Deep Etching ◽  
Quick Freezing ◽  
Acrosomal Reaction

Eggs of many species including those of echinoderms, amphibians and mammals exhibit an extensive extracellular matrix (ECM) that is important both in the reception of sperm and in providing a block to polyspermy after fertilization.In sea urchin eggs there are two distinctive coats, the vitelline layer which contains glycoprotein sperm receptors and the jelly layer that contains fucose sulfate glycoconjugates which trigger the acrosomal reaction and small peptides which act as chemoattractants for sperm. The vitelline layer (VL), as visualized by quick-freezing, deep-etching, and rotary-shadowing (QFDE-RS), is a fishnet-like structure, anchored to the plasma membrane by short posts. Orbiting above the VL are horizontal filaments which are thought to anchor the thicker jelly layer to the egg. Upon fertilization, the VL elevates and is transformed by cortical granule secretions into the fertilization envelope (FE). The rounded casts of microvilli in the VL are transformed into angular peaks and the envelope becomes coated inside and out with sheets of paracrystalline protein having a quasi-two dimensional crystalline structure.

1280: Increased Susceptibility to Genitovaginal Prolapse Associated with a Polymorphism in the Promoter of the Extracellular Matrix Protein LAMC1

2007 ◽  
Vol 177 (4S) ◽  
pp. 421-422
Author(s):  
Ganka Nikolova ◽  
Christian O. Twiss ◽  
Hane Lee ◽  
Nelson Stanley ◽  
Janet Sinsheimer ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Matrix Protein ◽  
Extracellular Matrix Protein ◽  
Increased Susceptibility

Protein gel blot analysis

2007 ◽  
Author(s):  
Dominique Loqué ◽  
Wolf Frommer
Keyword(s):  
Blot Analysis ◽  
Protein Gel

scholarly journals Homogenous overexpression of the extracellular matrix protein Netrin-1 in a hollow fiber bioreactor

2021 ◽  
Author(s):  
Aniel Moya-Torres ◽  
Monika Gupta ◽  
Fabian Heide ◽  
Natalie Krahn ◽  
Scott Legare ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Hollow Fiber ◽  
Mammalian Cells ◽  
Matrix Protein ◽  
Continuous Production ◽  
Extracellular Matrix Protein ◽  
Close Monitoring ◽  
High Quality ◽  
Biolayer Interferometry ◽  
Hollow Fiber Bioreactor

Abstract The production of recombinant proteins for functional and biophysical studies, especially in the field of structural determination, still represents a challenge as high quality and quantities are needed to adequately perform experiments. This is in part solved by optimizing protein constructs and expression conditions to maximize the yields in regular flask expression systems. Still, work flow and effort can be substantial with no guarantee to obtain improvements. This study presents a combination of workflows that can be used to dramatically increase protein production and improve processing results, specifically for the extracellular matrix protein Netrin-1. This proteoglycan is an axon guidance cue which interacts with various receptors to initiate downstream signaling cascades affecting cell differentiation, proliferation, metabolism, and survival. We were able to produce large glycoprotein quantities in mammalian cells, which were engineered for protein overexpression and secretion into the media using the controlled environment provided by a hollow fiber bioreactor. Close monitoring of the internal bioreactor conditions allowed for stable production over an extended period of time. In addition to this, Netrin-1 concentrations were monitored in expression media through biolayer interferometry which allowed us to increase Netrin-1 media concentrations tenfold over our current flask systems while preserving excellent protein quality and in solution behavior. Our particular combination of genetic engineering, cell culture system, protein purification, and biophysical characterization permitted us to establish an efficient and continuous production of high-quality protein suitable for structural biology studies that can be translated to various biological systems. Key points • Hollow fiber bioreactor produces substantial yields of homogenous Netrin-1 • Biolayer interferometry allows target protein quantitation in expression media • High production yields in the bioreactor do not impair Netrin-1 proteoglycan quality Graphical abstract

Direct interaction of the extracellular matrix protein DANCE with apolipoprotein(a) mediated by the kringle IV-type 2 domain

2002 ◽  
Vol 267 (4) ◽  
pp. 440-446 ◽  
Author(s):  
A. Kapetanopoulos ◽  
F. Fresser ◽  
G. Millonig ◽  
Y. Shaul ◽  
G. Baier ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Matrix Protein ◽  
Direct Interaction ◽  
Extracellular Matrix Protein ◽  
Apolipoprotein A
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