scholarly journals Menstrual-cycle-dependent expression of keratan sulphate in human endometrium

1990 ◽  
Vol 266 (3) ◽  
pp. 757-763 ◽  
Author(s):  
M E Hoadley ◽  
M W Seif ◽  
J D Aplin
Keyword(s):  
Monoclonal Antibody ◽  
Menstrual Cycle ◽  
Molecular Mass ◽  
Human Endometrium ◽  
Relative Molecular Mass ◽  
Keratan Sulphate ◽  
Sds Page ◽  
Immunochemical Methods ◽  
Cycle Dependent ◽  
Beta Galactosidase

Immunochemical methods have been used to detect and characterize two classes of polypeptide-associated keratan sulphate (KS) in epithelial secretions from human endometrium. Monoclonal antibody D9B1 binds to a hormonally regulated sialylated epitope associated with KS in a high relative molecular mass (250,000-350,000) component that bands as a doublet in SDS/PAGE. These KS chain(s) are sensitive to keratanase, endo-beta-galactosidase and N-glycanase. A second, more highly sulphated, type of KS is also present, that is resistant to all three enzymes. This can be detected using monoclonal antibody 5D4. It is present throughout the menstrual cycle and is associated principally with a component of Mr 140,000. Thus secretory KS contributes to the environment of the implanting embryo, may be used as a molecular index of endometrial function and could be important in the establishment of pregnancy.

A perichromosomal region contains proteins phosphorylated during mitosis in Xenopus laevis cells

1991 ◽  
Vol 98 (3) ◽  
pp. 309-315
Author(s):  
S.M. Dilworth
Keyword(s):  
Xenopus Laevis ◽  
Molecular Mass ◽  
Nuclear Protein ◽  
Antigenic Site ◽  
Relative Molecular Mass ◽  
Specific Expression ◽  
Storage Mechanism ◽  
Phosphorylated Form ◽  
Sds Page ◽  
Specific Manner

An antibody that recognizes the phosphorylated form of nucleoplasmin has identified another nuclear protein whose antigenic form is regulated in a mitosis-specific manner, with a dramatic increase in binding occurring in all mitotic cells. The protein is localised around the periphery of condensed chromosomes during mitosis in a manner analogous to another nucleoplasmin-related polypeptide NO38. Mitosis-specific expression of the antigenic site is dependent on phosphorylation of the polypeptide; binding of the antibody is dramatically reduced by prior incubation of the polypeptide with phosphatases. Migration on SDS-PAGE suggests that the protein has an exceptionally large relative molecular mass, in excess of 400,000. The probable mitosis-specific phosphorylation and location of this antigen suggests a subcellular storage mechanism for proteins during mitosis.

scholarly journals PROTEINS OF PAROTOID GLAND SECRETIONS FROM TOADS OF THE GENUS BUFO

2000 ◽  
pp. 1-7 ◽  
Author(s):  
David Perry
Keyword(s):  
Amino Acid ◽  
Molecular Mass ◽  
Mass Range ◽  
Relative Molecular Mass ◽  
Terminal Amino Acid ◽  
Banding Patterns ◽  
Freeze Dried ◽  
Sds Page ◽  
Terminal Amino ◽  
Terminal Amino Acid Sequence

Freeze-dried parotoid gland secretions from toads of the genus Bufo contained large proportions of protein (25-35% by weight). SDS-PAGE suggested that secretions from several species of Bufo contained mixtures of proteins in the relative molecular mass range of approximately 12 - 200 kDa, which exhibited markedly different banding patterns from species to species. These proteins were presumably not discovered before because the previous extraction procedures used with these secretions were designed to examine low molecular mass compounds and would denature the proteins. SDS-PAGE of secretions from B. mauritanicus and B. calamita are shown here. The N-terminal amino acid sequence of one of the bands (approx. 58 kDa) of B. mauritanicus was found to be LPIPAFPGLDHGF and of a B. calamita band (30.5 kDa) was VQVFGLQKEA. No significant similarities to these two sequences and to three separate but partial N-terminal sequences obtained from these species were found in genetic databases.

scholarly journals The Normal Human Endometrium but not Myometrium Presents Menstrual Cycle-Dependent Fluctuations in the Immunoexpression of DNA Fragmentation Factor 40, DNA Fragmentation Factor 45, and B-cell Lymphoma 2 Protein.

2017 ◽  
Author(s):  
Tomasz Banas ◽  
Kazimierz Pitynski ◽  
Krzysztof Okon ◽  
Marcin Mikos ◽  
Joanna Bonior ◽  
...  
Keyword(s):  
Menstrual Cycle ◽  
B Cell ◽  
Dna Fragmentation ◽  
Cell Lymphoma ◽  
B Cell Lymphoma ◽  
Human Endometrium ◽  
Proliferative Endometrium ◽  
Normal Human ◽  
Cycle Dependent ◽  
Secretory Endometrium

Abstract: DNA fragmentation factors 40 and 45 (DFF40 and DFF45) and B-cell lymphoma 2 (Bcl-2) expression were evaluated in the normal human endometrium and myometrium. DFF40, DFF45, and Bcl-2 expression was assessed via immunohistochemistry in the proliferative, secretory, and atrophic endometrium and myometrium collected postmenopausally and premenopausally during the proliferative and secretory phases of the menstrual cycle. The endometrium showed significantly higher DFF40 and DFF45 expression than that in the uterine myometrium; compared to the stroma, endometrial glands showed the highest expression in pre- and postmenopausal specimens. Glandular expression of DFF45 was dependent on the menstrual cycle, reaching its highest level in the secretory endometrium. The glandular expression of DFF40 and DFF45 was significantly lower in postmenopausal specimens than that in premenopausal tissue. No cycle-dependent changes were reported for stromal or myometrial DFF40 or DFF45 expression. Compared to the endometrial stroma and myometrium, Bcl-2 showed the highest expression in the glandular proliferative endometrium and the lowest expression in the stromal secretory endometrium and myometrium during the secretory phase of the cycle. DFF45 and Bcl-2 showed menstrual cycle-dependent expression, which was limited to the glandular layer of the endometrium.

scholarly journals Characterization of proteoglycans isolated from associative extracts of human articular cartilage

1993 ◽  
Vol 293 (1) ◽  
pp. 165-172 ◽  
Author(s):  
V Vilím ◽  
A J Fosang
Keyword(s):  
Articular Cartilage ◽  
Molecular Mass ◽  
Core Protein ◽  
Chondroitin Sulphate ◽  
Gel Chromatography ◽  
Human Articular Cartilage ◽  
Keratan Sulphate ◽  
Core Proteins ◽  
Sds Page

Approx. 10% of the total proteoglycan content of normal young human articular cartilage was extracted under associative conditions with Dulbecco's PBS. Proteoglycans isolated from the extract by Q-Sepharose chromatography were separated by gel chromatography and characterized by gradient gel SDS/PAGE and immunoblotting. Three species of small proteoglycans, two main populations of aggrecan and a population of its smaller fragments were identified. The major populations of aggrecan contained chondroitin sulphate chains, all or part of the N-terminal G1 and G2 domains and, therefore, intact keratan sulphate domains. The larger population was estimated by gradient SDS/PAGE to have a molecular mass of approx. 600 kDa or greater. The second population had an apparent molecular mass of approx. 300-600 kDa. Core proteins derived from these populations of proteoglycans separated on SDS/PAGE into several clusters of bands in the range from 120 to approx. 360 kDa. The extract further contained smaller fragments which lacked chondroitin sulphate but reacted with antibodies against keratan sulphate, and against epitopes present in the G2 domain of aggrecan. The presence of the G2 domain in a broad range of populations of decreasing size indicated extensive cleavage of the aggrecan core protein within its chondroitin sulphate domain. These findings suggest that fragmentation of aggrecan probably occurs in vivo in normal articular cartilage of young individuals. Associative extracts also contained decorin, biglycan and fibromodulin. These were resolved from aggrecan by gel chromatography and identified by immunodetection.

scholarly journals Human liver N-acetylgalactosamine 6-sulphatase. Purification and characterization

1991 ◽  
Vol 279 (2) ◽  
pp. 515-520 ◽  
Author(s):  
J Bielicki ◽  
J J Hopwood
Keyword(s):  
Molecular Mass ◽  
Gel Filtration ◽  
Chloride Ions ◽  
Lysosomal Degradation ◽  
Purification And Characterization ◽  
Keratan Sulphate ◽  
Native Molecular Mass ◽  
Sds Page ◽  
Molecular Masses ◽  
Column Procedure

Human N-acetylgalactosamine 6-sulphatase (EC 3.1.6.14), which is involved in the lysosomal degradation of the glycosaminoglycans keratan sulphate and chondroitin 6-sulphate, was purified more than 130,000-fold in 2.8% yield from liver by an eight-step column procedure. One major form was identified with a pI of 5.7 and a native molecular mass of 62 kDa by gel filtration. When analysed by SDS/PAGE, dithioerythritol-reduced enzyme contained polypeptides of molecular masses 57 kDa, 39 kDa and 19 kDa, whereas non-reduced enzyme contained a major polypeptide of molecular mass 70 kDa. It is proposed that active enzyme contains either the 57 kDa polypeptide or disulphide-linked 39 kDa and 19 kDa polypeptides. Minor amounts of other enzyme forms separated during the chromatofocusing step and the Blue A-agarose step were not further characterized. Purified N-acetylgalactosamine 6-sulphatase was inactive towards 4-methylumbelliferyl sulphate, but was active, with pH optima of 3.5-4.0, towards 6-sulphated oligosaccharide substrates. Km values of 12.5 and 50 microM and Vmax. values of 1.5 and 0.09 mumol/min per mg were determined with oligosaccharide substrates derived from chondroitin 6-sulphate and keratan sulphate respectively. Sulphate, phosphate and chloride ions were inhibitors of enzyme activity towards both substrates, with 50 microM-Na2SO4 giving 50% inhibition towards the chondroitin 6-sulphate trisaccharide substrate.

scholarly journals Vinculin is a major platelet protein that undergoes Ca2+-dependent tyrosine phosphorylation

1993 ◽  
Vol 294 (3) ◽  
pp. 675-680 ◽  
Author(s):  
J G Vostal ◽  
N R Shulman
Keyword(s):  
Monoclonal Antibody ◽  
Molecular Mass ◽  
Tyrosine Phosphorylation ◽  
Platelet Activation ◽  
Soluble Fraction ◽  
Phosphorylated Protein ◽  
Platelet Protein ◽  
Activated Platelets ◽  
Sds Page ◽  
Tyrosine Phosphorylated Protein

When intracellular Ca2+ pools are released during platelet stimulation by thrombin, elevation of platelet cytosolic Ca2+ concentration induces tyrosine phosphorylation of a 130 kDa protein, and refilling the pools mediates dephosphorylation of this protein [Vostal, Jackson and Shulman (1991) J. Biol. Chem. 266, 16911-16916]. In the present work the 130 kDa protein was identified as vinculin by the following criteria. (1) It is detected on protein immunoblots of thrombin-activated platelets by both monoclonal anti-phosphotyrosine and anti-vinculin antibodies. (2) Removal of N-linked sugars with peptide-N-glycosidase or reduction did not change the molecular mass of vinculin or of the 130 kDa protein on SDS/PAGE. (3) The 130 kDa tyrosine-phosphorylated protein associates with Triton-soluble fraction of platelets as does vinculin. (4) The 130 kDa protein immunoprecipitated by anti-vinculin monoclonal antibody reacts with anti-phosphotyrosine antibody; when immunoprecipitated by anti-phosphotyrosine antibody it reacts with anti-vinculin antibody. (5) The 130 kDa tyrosine-phosphorylated protein and vinculin focus isoelectrically at pI 5.4-5.8. Our finding that vinculin is a major platelet protein that undergoes Ca(2+)-dependent tyrosine phosphorylation during platelet activation may provide clues to the function of this protein.

scholarly journals Uji Aktivitas Lektin Biji Kebiul terhadap Kecepatan Penggumpalan Sel Darah Merah Manusia dalam Kondisi Patologis dan Implementasinya sebagai Modul Pembelajaran Kimia

2018 ◽  
Vol 2 (2) ◽  
Author(s):  
Yeni Trianah
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Molecular Mass ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Seed Extract ◽  
Relative Molecular Mass ◽  
Human Red Blood Cells ◽  
Sds Page ◽  
Post Test

ABSTRACT[Lectin activities of kebiul seeds to the red blood cell agglutination speed in pathological condition and its implementation as a chemical learning module]. The purposes of this research were to determine: 1) the relative moleculer mass protein that behaves as a lectin in the seed extract kebiul , 2) the velocity of red blood cell clumping influenced by seed lectin kebiul, 3) know the difference student results on protein taught modules and a without modules taught at the College of Teacher Training and Education Teachers Association of the Republic of Indonesia (STKIP-PGRI) Lubuklinggau. Extraction of seeds Kebiul carried out in the a cold buffer solution with pH 7.4 and plus 60% saturated ammonium sulfat (salting out method), and made in four concentrations, namely : 2%, 4%, 6% and 8%. Then tested the activity of seed lectin kebiul to speed clotting of human red blood cells in pathological conditions. To determine the relative molecular mass protein that behaves as a lectin in the seed extract kebiul SDS PAGE electrophoresis performed 1-D. The experiment were then implemented on the material of protein biochemistry using modules. The results of the research showed that on the concentration of 8% the velocity of the clumping of a human red blood cells hypertension most of the highest, the relative molecular mass which behaves as a lectin protein electrophoresis results of 1-D SDS PAGE obtained by a three protein bands in the range of moleculer weights 80, 128 and 144 kDa. The Results of the implementation of the experimental class showed an average `post test value was 95 and the post test control class 69.41. There are differences in students' learning about protein for students who are taught by module and who are taught without module.Keywords: Lectin; agglutination; blood; 1-D SDS PAGE; learning outcomes.

scholarly journals Uji Aktivitas Lektin Biji Kebiul terhadap Kecepatan Penggumpalan Sel Darah Merah Manusia dalam Kondisi Patologis dan Implementasinya sebagai Modul Pembelajaran Kimia

2018 ◽  
Vol 2 (2) ◽  
pp. 214-221
Author(s):  
Yeni Trianah
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Molecular Mass ◽  
Red Blood Cell ◽  
Blood Cells ◽  
Seed Extract ◽  
Relative Molecular Mass ◽  
Human Red Blood Cells ◽  
Sds Page ◽  
Post Test

ABSTRACT[Lectin activities of kebiul seeds to the red blood cell agglutination speed in pathological condition and its implementation as a chemical learning module]. The purposes of this research were to determine: 1) the relative moleculer mass protein that behaves as a lectin in the seed extract kebiul , 2) the velocity of red blood cell clumping influenced by seed lectin kebiul, 3) know the difference student results on protein taught modules and a without modules taught at the College of Teacher Training and Education Teachers Association of the Republic of Indonesia (STKIP-PGRI) Lubuklinggau. Extraction of seeds Kebiul carried out in the a cold buffer solution with pH 7.4 and plus 60% saturated ammonium sulfat (salting out method), and made in four concentrations, namely : 2%, 4%, 6% and 8%. Then tested the activity of seed lectin kebiul to speed clotting of human red blood cells in pathological conditions. To determine the relative molecular mass protein that behaves as a lectin in the seed extract kebiul SDS PAGE electrophoresis performed 1-D. The experiment were then implemented on the material of protein biochemistry using modules. The results of the research showed that on the concentration of 8% the velocity of the clumping of a human red blood cells hypertension most of the highest, the relative molecular mass which behaves as a lectin protein electrophoresis results of 1-D SDS PAGE obtained by a three protein bands in the range of moleculer weights 80, 128 and 144 kDa. The Results of the implementation of the experimental class showed an average `post test value was 95 and the post test control class 69.41. There are differences in students' learning about protein for students who are taught by module and who are taught without module.Keywords: Lectin; agglutination; blood; 1-D SDS PAGE; learning outcomes.

scholarly journals Activation of the cell wall degrading protease, lysin, during sexual signalling in Chlamydomonas: the enzyme is stored as an inactive, higher relative molecular mass precursor in the periplasm.

1989 ◽  
Vol 108 (1) ◽  
pp. 199-207 ◽  
Author(s):  
M J Buchanan ◽  
S H Imam ◽  
W A Eskue ◽  
W J Snell
Keyword(s):  
Cell Wall ◽  
Molecular Mass ◽  
Mating Type ◽  
Gradient Centrifugation ◽  
Cell Contact ◽  
Relative Molecular Mass ◽  
Adhesive Interaction ◽  
Sds Page ◽  
Sexual Signalling ◽  
A Cell

During the mating reaction in Chlamydomonas reinhardtii mating type plus and mating type minus gametes adhere to each other via adhesion molecules on their flagellar surfaces. This adhesive interaction induces a sexual signal leading to release of a cell wall degrading enzyme, lysin, that causes wall release and degradation. In this article, we describe the preparation of a polyclonal antibody against the 60,000-Mr lysin polypeptide excised from SDS-PAGE gels. After absorption of the IgG with cell walls to remove antibodies against a carbohydrate epitope common to several Chlamydomonas glycoproteins, the immune IgG reacted with the 60,000-Mr polypeptide, and a 47,000-Mr species that we show here was immunologically cross-reactive with the 60,000-Mr molecule. By use of several fractionation methods including ion exchange and molecular sieve chromatography, sucrose gradient centrifugation, and affinity chromatography, we showed that the 60,000-Mr antigen copurified with lysin activity, thereby demonstrating that the antibody was indeed directed against the enzyme. Immunoblot experiments on suspensions of nonmating and mating gametes showed that the 60,000-Mr antigen was missing in the nonmating gametes. Instead, they contained a 62,000-Mr antigen that was not present in suspensions of mating gametes that had undergone sexual signalling. Furthermore, nonmating gametes whose walls were removed with exogenously added lysin did not contain either form of the antigen. We also found that the 62,000-Mr form of the antigen, which could be released from gametes by freeze-thawing, did not have wall degrading activity. These results indicate that lysin in gametes is stored in the periplasm as a higher relative molecular mass, inactive precursor and also that sexual signalling induces conversion of this molecule to a lower relative molecular mass, active enzyme. This may be a novel example of processing of an extracellular protease induced by cell contact.

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