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 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 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.

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 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.

Detection of both Type 1 and Type 2 Plasminogen Activator Inhibitors in Human Monocytes

1990 ◽  
Vol 63 (01) ◽  
pp. 067-071 ◽  
Author(s):  
Joan C Castellote ◽  
Enric Grau ◽  
Maria A Linde ◽  
Nuria Pujol-Moix ◽  
Miquel LI Rutllant
Keyword(s):  
Molecular Mass ◽  
Plasminogen Activator ◽  
Human Peripheral Blood ◽  
Direct Interaction ◽  
Apparent Molecular Weight ◽  
Fibrinolytic System ◽  
Human Monocytes ◽  
Blood Monocytes ◽  
Single Chain ◽  
Sds Page

SummaryIncreasing evidence suggests the involvement of leukocytes in the fibrinolytic system. Monocytes secrete pro-urokinase (Grau, Thromb Res 1989; 53: 145) and it has been shown that these cells have specific receptors for urokinase and plasminogen (Miles, Thromb Haemostas 1987; 58: 936). The aim of this study was to analyse the presence of plasminogen activator inhibitor(s) in platelet-free suspensions of human peripheral blood monocytes and polymorphonuclear leukocytes (PMN). SDS-PAGE and reverse fibrin autography showed an inhibitory band of 50 kDa in the monocyte extracts (Triton X-100) but not in the PMN extracts. Urokinase (u-PA) was mixed with increasing amounts of monocyte extract for 10 min and the mixtures were added to 125Ifibrin coated wells containing plasminogen. A dose-dependent decrease in the u-PA fibrinolytic activity was observed. The amount of inhibition increased when the monocyte releasates were preincubated with u-PA (40% inhibition after 5 min preincubation and 80% after 15 min), indicating a direct interaction between this activator and an inhibitor(s). After SDS-PAGE of monocyte extracts, immunoblotting and peroxidase staining identified both PAI1 and PAI2, with an apparent molecular weight of 47-50 kDa. Monocyte-associated PAI1 formed complexes with single chain t-PA with a molecular mass 50 kDa higher than the molecular mass of the free PAI1. However, a significant amount of PAI remained unbound to t-PA. This inactive PAI1 could have come from a rapid inactivation of the primary active PAI1. These PAI1 and PAI2 detected in human monocytes may be transcendent in the regulation of the fibrinolytic system.

Extraction of polyethylene glycols with dicarbolide solutions in nitrobenzene

1990 ◽  
Vol 55 (8) ◽  
pp. 1959-1967 ◽  
Author(s):  
Petr Vaňura ◽  
Pavel Selucký
Keyword(s):  
Polyethylene Glycol ◽  
Molecular Mass ◽  
Extraction Constant ◽  
Metal Extraction ◽  
Polyethylene Glycols ◽  
Relative Molecular Mass ◽  
Published Data ◽  
Average Molecular Mass ◽  
Peg 400 ◽  
Extraction Constants

The extraction of polyethylene glycol of average molecular mass 400 (PEG 400) with dicarbolide solution in nitrobenzene and of longer-chain polyethylene glycol, of average molecular mass 1 500 (PEG 1 500), with chlorinated dicarbolide solution in nitrobenzene was studied. During the extraction of PEG 400, the polyethylene glycol solvates the Horg+ ion in the organic phase giving rise to the HLorg+ species (L is polyethylene glycol). The obtained value of the extraction constant Kex(HLorg+) = 933 is consistent with published data of metal extraction. Extraction of PEG 1 500 was treated applying the simplified assumption that the thermodynamic behaviour of PEG 1 500 is the same as that of n molecules of polyethylene glycol with relative molecular mass 1 500/n, each solvating one cation. For this model, the value of n = 3.2 ± 1.1 and the values of the extraction constants of the HL1/n,org+ and HL2/n,org+ species were obtained by using the adapted program LETAGROP. This value of n is consistent with published extraction data in the presence of polyethylene glycol with a relative molecular mass from 200 to 1 000.

scholarly journals Identification and characterization of Sulfolobus solfataricusD-gluconate dehydratase: a key enzyme in the non-phosphorylated Entner–Doudoroff pathway

2005 ◽  
Vol 387 (1) ◽  
pp. 271-280 ◽  
Author(s):  
Seonghun KIM ◽  
Sun Bok LEE
Keyword(s):  
Molecular Mass ◽  
Gel Filtration ◽  
Maximal Activity ◽  
Genome Database ◽  
Sds Page ◽  
Key Enzyme ◽  
Bivalent Metal Ions ◽  
Ammonium Sulphate Fractionation ◽  
Identification And Characterization

The extremely thermoacidophilic archaeon Sulfolobus solfataricus utilizes D-glucose as a sole carbon and energy source through the non-phosphorylated Entner–Doudoroff pathway. It has been suggested that this micro-organism metabolizes D-gluconate, the oxidized form of D-glucose, to pyruvate and D-glyceraldehyde by using two unique enzymes, D-gluconate dehydratase and 2-keto-3-deoxy-D-gluconate aldolase. In the present study, we report the purification and characterization of D-gluconate dehydratase from S. solfataricus, which catalyses the conversion of D-gluconate into 2-keto-3-deoxy-D-gluconate. D-Gluconate dehydratase was purified 400-fold from extracts of S. solfataricus by ammonium sulphate fractionation and chromatography on DEAE-Sepharose, Q-Sepharose, phenyl-Sepharose and Mono Q. The native protein showed a molecular mass of 350 kDa by gel filtration, whereas SDS/PAGE analysis provided a molecular mass of 44 kDa, indicating that D-gluconate dehydratase is an octameric protein. The enzyme showed maximal activity at temperatures between 80 and 90 °C and pH values between 6.5 and 7.5, and a half-life of 40 min at 100 °C. Bivalent metal ions such as Co2+, Mg2+, Mn2+ and Ni2+ activated, whereas EDTA inhibited the enzyme. A metal analysis of the purified protein revealed the presence of one Co2+ ion per enzyme monomer. Of the 22 aldonic acids tested, only D-gluconate served as a substrate, with Km=0.45 mM and Vmax=0.15 unit/mg of enzyme. From N-terminal sequences of the purified enzyme, it was found that the gene product of SSO3198 in the S. solfataricus genome database corresponded to D-gluconate dehydratase (gnaD). We also found that the D-gluconate dehydratase of S. solfataricus is a phosphoprotein and that its catalytic activity is regulated by a phosphorylation–dephosphorylation mechanism. This is the first report on biochemical and genetic characterization of D-gluconate dehydratase involved in the non-phosphorylated Entner–Doudoroff pathway.

Towards a molecular description of cell transformation by avian sarcoma virus

1980 ◽  
Vol 210 (1180) ◽  
pp. 387-396 ◽  
Keyword(s):  
Protein Kinase ◽  
Protein Phosphorylation ◽  
Molecular Mass ◽  
Cellular Protein ◽  
Transformed Cells ◽  
Relative Molecular Mass ◽  
Protein Kinase Activity ◽  
Avian Sarcoma Virus ◽  
Sarcoma Virus ◽  
Avian Sarcoma

The avian sarcoma virus transforming gene product has been identified and partially purified from extracts of transformed cells. It is a phosphoprotein with a relative molecular mass of 60 000 (pp60 src ) with two major sites of phosphorylation. pp60 src appears to be a cyclic-AMP-independent protein kinase as judged by protein phosphorylation with partly purified fractions. The specificity of the phosphorylation observed was judged by inhibition with anti-pp60 src IgG but not by normal IgG and by the fact that the protein kinase activity isolated from ts transformation-mutant infected cells was more thermolabile than that from wild-type transformed cells, thus showing more directly the origin of the enzymic activity. A cellular protein substrate of pp60 src has been identified as a 34000 molecular mass protein. These data together suggest that protein phosphorylation by pp60 src may be a function of the molecule that plays a major role in transformation.

Sign in / Sign up

Export Citation Format

Share Document