scholarly journals Isolation of a novel protein from the outer layer of the vitelline membrane

1992 ◽  
Vol 286 (1) ◽  
pp. 17-22 ◽  
Author(s):  
S Kido ◽  
A Morimoto ◽  
F Kim ◽  
Y Doi

The outer layer of the vitelline membrane from hen egg yolk consists of ovomucin, vitelline membrane outer layer protein I (VMOI) and lysozyme. Here we report the occurrence of a further basic protein (pI 11.5) in the outer layer, which was designated as vitelline membrane outer layer protein II (VMOII). It was dissociated from the outer layer in a 10% (w/v) NaCl solution and purified to homogeneity by ion-exchange chromatography. VMOII is a simple protein with a molecular mass of 6000 Da, as determined by sedimentation equilibrium analysis. The amino acid composition of VMOII was characterized by the absence of Met and high contents of cystine (half) (14%) and basic amino acids (6% Arg, 6% Lys and 3% His). Analysis of carboxymethylated VMOII indicated that all cysteine residues were involved in disulphide bonding, which appears to facilitate the binding of SDS to the protein. Sequence comparison of the N-terminal 20 residues revealed no identity with other known proteins. VMOII contained a small amount of alpha-helix and was quite resistant to heat denaturation.

1969 ◽  
Vol 22 (3) ◽  
pp. 653 ◽  
Author(s):  
Joan M Bain ◽  
Janice M Hall

Stages in the development of the outer layer of the vitelline membrane of a hen's egg have been observed in an egg found in the infundibulum of a sacrificed White Leghorn hen. Tissue from the infundibulum and the underlying egg yolk material was taken at increasing distances from the upper end of the egg and the relationship between the secretory cells of the infundibulum and the vitelline mem-brane observed. The structure of the vitelline membrane in ova just liberated from the ovary and not yet in the oviduct and that of the vitelline membrane in new-laid eggs from other White Leghorn hens were observed for comparison.


2017 ◽  
Author(s):  
Jamil Momand ◽  
Patrycja Magdziarz ◽  
You Feng ◽  
Dianlu Jiang ◽  
Elizabeth Parga ◽  
...  

ABSTRACTt-Darpp is a protein encoded by the PPP1R1B gene and is expressed in breast, colon, esophageal, gastric, and prostate cancers, as well as in normal adult brain striatal cells. Overexpression of t-Darpp in cultured cells leads to increased protein kinase A activity and increased phosphorylation of AKT (protein kinase B). In HER2+ breast cancer cells t-Darpp confers resistance to the chemotherapeutic agent trastuzumab. To shed light on t-Darpp function, we studied its secondary structure, oligomerization status, metal-binding properties, and phosphorylation by cyclin dependent kinases 1 and 5. t-Darpp exhibits 12% alpha helix, 29% beta strand, 24% beta turn and 35% random coil structures. t-Darpp binds to calcium, but not to other metals commonly found in biological systems. The T39 site, critical for t-Darpp activation of the AKT signaling pathway, is a substrate for phosphorylation by cyclin-dependent kinase 1 (CDK1) and cyclin-dependent kinase 5 (CDK5). Gel filtration chromatography, sedimentation equilibrium analysis, blue native gel electrophoresis, and glutaraldehyde-mediated crosslinking experiments demonstrate that the majority of t-Darpp exists as a monomer, but forms low levels (< 3%) of hetero-oligomers with its longer isoform Darpp-32. t-Darpp has a large Stokes radius of 4.4 nm relative to its mass of 19 kDa, indicating that it has an elongated structure.


2006 ◽  
Vol 69 (5) ◽  
pp. 1012-1016 ◽  
Author(s):  
TOSHIYUKI MURASE ◽  
KAZUHIKO FUJIMOTO ◽  
RUI NAKAYAMA ◽  
KOICHI OTSUKI

The invasive ability of Salmonella enterica serovars Enteritidis, Infantis, and Montevideo in eggs was examined. Strains of these serovars originating from egg contents, laying chicken houses, and human patients were experimentally inoculated (0.1-ml dose containing 78 to 178 cells) onto the vitelline membrane of eggs collected from specific-pathogen-free chickens and incubated at 25°C. The test strains were detected in 25 of 138 yolk contents by day 6, indicating the penetration of Salmonella organisms through the vitelline membrane. There were no significant differences in overall rates of penetration between serovars. The organisms were also detected in the albumen from 125 of 138 eggs tested by day 6. Growth to more than 106 CFU/ml was observed in 48 of the 125 albumen samples. An inoculum of 1,000 Salmonella cells was added to 15 ml of albumen at the edge of a petri plate. A 10-mm-diameter cylindrical well, the bottom of which was sealed with a polycarbonate membrane with 3.0-μm pores, was filled with egg yolk and placed into the albumen at the center of the dish, which was maintained at 25°C. Experiments were performed in triplicate with each strain. Salmonella organisms in all the albumen samples were detected by day 11. However, motility of the organisms toward the yolk was observed in only two dishes inoculated with the Salmonella Enteritidis strain from a human patient and in one dish inoculated with the Salmonella Infantis strain from liquid egg. The albumen samples obtained from the dishes inoculated with the Salmonella Enteritidis strain had high numbers of bacteria (&gt;108 CFU/ml). The present study suggests that Salmonella organisms in egg albumen are unlikely to actively move toward the yolk, although deposition on or near the vitelline membrane can be advantageous for proliferation.


1886 ◽  
Vol s2-26 (102) ◽  
pp. 157-174
Author(s):  
WALTER HEAPE

The membranes surrounding the ripe ovarian, ovum are two: (1) a single outer, thick, zona radiata, with a granular peripheral and a transparent inner portion, pierced radially by fine canals through which nutriment is obtained by the ovum from, the follicular cells (of the discus proligerus) immediately in contact with the zona: (2) an inner very delicate vitelline membrane which closely covers the ovum itself; and between these membranes is a space, the circum-vitelline space. The confirmation of Reichert's (No. 18), Meyer's (No. 17), and van Beneden's (No. 4) observations as to the presence of the inner delicate vitelline membrane appears of some interest as many embryologists are still sceptical of its existence, while the relation of the follicular cells with the radial canals of the zona supports the view as to the source of the nutriment of the ovarian ovum. On the other hand the fact that nothing was seen comparable to a micropyle in the zona, such as M. Barry (No. 3), and Meissner (No. 16), described, nor any follicular cells within the zona such as Lindgren (No. 15), von Sehlen (No. 21), and Virchow (No. 22), have observed, is some further proof that the conditions of the material investigated by these authors was abnormal. The yolk contained within the ovum, which is of two kinds: viz. (1) homogeneous vesicular bodies, (2) minute highly refractile granules, is contained within the meshes of a protoplasmic reticulum; it is dense and contains no large globules such as Beneden (Nos. 6 and 7) describes in theBat's ova. The rounded or oval nucleus contains a single centrally placed nucleolus and a variable number of smaller or larger granules, which may possibly be considered as nucleolar material. During maturation the vitellus becomes divided into a medullary granular, and a cortical non-granular portion, the circum-vitelline space between the zona radiata and the vitelline membrane is enlarged, while the vitellus itself contracts away from the vitelline membrane excepting (1) here and there where pseudopodia-like processes connect the two, and (2) at one spot where the polar bodies are formed. At this latter place two polar bodies may be seen in the specimen figured, outside the vitelline membrane, whilst the nucleus remains as the female pronucleus lying in the peripheral portion of the ovum. Finally, the vitellus again expands and the nucleus retires to the centre of the ovum and is no longer to be seen. Assuming that these observations are correct, Beneden's description of the ejection of the vesicle to form the polar bodies and the subsequent non-nucleated condition of the ovum must be considered erroneous. Impregnation appears to be effected by a single spermatazoon, although a considerable number of spermatazoa find their way through the zona and may be seen lying passively in the circumvitelline space. The segmentation occurs while the ovum travels down the Fallopian tube. Two and then four segments are formed, after which the course of segmentation is irregular. The segments themselves are of irregular size and do not appear to be divisible into two kinds (epiblastic and hypoblastic) as Beneden describes. After its entrance into the uterus, a division of the segments into an outer hyaline layer and inner deeply granular mass takes place, and I would suggest the hypothesis that the vitelline matter which was originally contained in all segments alike has been transmitted from the outer segments to the segments lying in the interior of the ovum, in order that the former segments may the more readily and actively multiply and flatten out to form the wall of the blastodermic vesicle. The epiblast of the vesicle and of the embryo is derived from the whole of the outer layer and by far the largest proportion of the inner mass of segments. The hypoblast is derived from the small remaining portion of the inner mass and the mesablast, subsequently, from both epiblast and hypoblast layers. This being the case, the division of the segmentation spheres, by Beneden, into epiblast and hypoblast spheres from the time when the first two segments were formed, is incorrect; and at the same time the theory of a comparison of the metagastrula stage with the gastrula of other animals is likewise untenable.


1995 ◽  
Vol 117 (6) ◽  
pp. 1183-1191 ◽  
Author(s):  
Shoko Kido ◽  
Yukio Doi ◽  
Fimi Kim ◽  
Emi Morishita ◽  
Hiroshi Narita ◽  
...  

1984 ◽  
Vol 98 (1) ◽  
pp. 214-221 ◽  
Author(s):  
P C Tseng ◽  
M S Runge ◽  
J A Cooper ◽  
R C Williams ◽  
T D Pollard

Acanthamoebe profilin has a native molecular weight of 11,700 as measured by sedimentation equilibrium ultracentrifugation and an extinction coefficient at 280 nm of 1.4 X 10(4) M-1cm-1. Rabbit antibodies against Acanthamoeba profilin react only with the 11,700 Mr polypeptide among all other ameba polypeptides separated by electrophoresis. These antibodies react with a 11,700 Mr polypeptide in Physarum but not with any proteins of Dictyostelium or Naeglaria. Antibody-binding assays indicate that approximately 2% of the ameba protein is profilin and that the concentration of profilin is approximately 100 mumol/liter cells. During ion exchange chromatography of soluble extracts of Acanthamoeba on DEAE-cellulose, the immunoreactive profilin splits into two fractions: an unbound fraction previously identified by Reichstein and Korn (1979, J. Biol. Chem., 254:6174-6179) and a tightly bound fraction. Purified profilin from the two fractions is identical by all criteria tested. The tightly bound fraction is likely to be attached indirectly to the DEAE, perhaps by association with actin. By fluorescent antibody staining, profilin is distributed uniformly throughout the cytoplasmic matrix of Acanthamoeba. In 50 mM KCl, high concentrations of Acanthamoeba profilin inhibit the elongation rate of muscle actin filaments measured directly by electron microscopy, but the effect is minimal in KCl with 2 MgCl2. By using the fluorescence change of pyrene-labeled Acanthamoeba actin to assay for polymerization, we confirmed our earlier observation (Tseng, P. C.-H., and T. D. Pollard, 1982, J. Cell Biol. 94:213-218) that Acanthamoeba profilin inhibits nucleation much more strongly than elongation under physiological conditions.


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