Heterogeneity of Human Biliary Mucin: Functional Implications

1994 ◽  
Vol 86 (1) ◽  
pp. 75-82 ◽  
Author(s):  
J. Henriëtte Klinkspoor ◽  
Michel J. A. van Wijland ◽  
Carolien A. M. Koeleman ◽  
Willem van Dijk ◽  
Guido N. J. Tytgat ◽  
...  
Keyword(s):  
Cholesterol Gallstone ◽  
Lectin Binding ◽  
Gallstone Disease ◽  
Functional Characterization ◽  
The Other ◽  
Binding Studies ◽  
Human Gallbladder ◽  
Patient Heterogeneity ◽  
Model Bile

1. Human gallbladder mucin has been implicated as playing a role in the pathogenesis of gallstones. In previous studies no differences have been found in the content or composition of mucins derived from control bile or cholesterol gallstone bile. Until now, no differences were also found between these two groups of mucins with regard to their ability to cause cholesterol nucleation. In the accompanying paper we have reported that there is a strong heterogeneity of gallbladder mucins derived from individual patients (M. J. A. van Wijland, J. H. Klinkspoor, L. Th. de Wit, R. P. J. Oude Elferink, G. N. J. Tytgat and A. K. Groen, Clin Sci 1994; 86: 67–74). In the present study we further investigated a possible patient to patient heterogeneity of mucin by means of immunological and functional characterization of mucins isolated from hepatic bile of six different patients with gallstones. 2. Considerable heterogeneity was found. Two of the mucins barely reacted with a polyclonal anti-mucin antibody, whereas the other four mucins reacted very strongly. Lectin-binding studies indicated that the glycans of these two mucins expressed less D-N-acetylgalactosamine residues than the other four mucins. This was confirmed by analyses of the glycan compositions. These studies furthermore indicated that the glycans were of the O-linked type, contained α-D-N-acetylglucosamine and were fucosylated, sialy-lated and sulphated to different extents. Except for a strong heterogeneity in the sugar composition of the mucins, heterogeneity was also found in the biological activity of the mucins. The two immunologically diverging mucins nucleated cholesterol from model bile 1–2 days earlier and also caused an almost threefold more rapid rupture of cholesterol/phospholipid vesicles than the other mucins. 3. We conclude that considerable differences exist between mucins derived from individual patients and that the heterogeneity in glycan composition might play a role in the pathogenesis of gallstone disease.

scholarly journals Fibronectin in human gallbladder bile: cholesterol pronucleating and/or mucin "link" protein?

1994 ◽  
Vol 267 (3) ◽  
pp. G393-G400 ◽  
Author(s):  
J. F. Miquel ◽  
C. Von Ritter ◽  
R. Del Pozo ◽  
V. Lange ◽  
D. Jungst ◽  
...  
Keyword(s):  
Cholesterol Gallstone ◽  
Gallstone Disease ◽  
Gallstone Formation ◽  
Gallbladder Bile ◽  
Crystal Formation ◽  
Nucleation Time ◽  
Human Gallbladder ◽  
Link Protein ◽  
Wide Range ◽  
Model Bile

Some biliary proteins (pronucleators) seem to be essential factors for cholesterol crystal formation and crystal growth in bile. A recent study suggests that fibronectin is such a pronucleator in bile. Fibronectin also seems to closely interact with intestinal mucin. Since biliary mucin plays an important role in gallstone formation, such an interaction in bile may be of relevance in cholesterol gallstone formation. To more clearly elucidate the role of fibronectin in cholesterol gallstone disease, we measured the concentration of fibronectin in native bile of cholesterol gallstone patients and checked its influence on the cholesterol nucleation time of model bile. We further looked for a molecular interaction between biliary fibronectin and gallbladder mucin. We found that fibronectin is present in gallbladder bile of gallstone patients in low concentrations (2.6 +/- 1.2 micrograms/ml). Bile fibronectin did not interact with gallbladder mucin. Moreover, in a wide range of concentrations fibronectin had no influence on the nucleation time of model bile. We conclude that fibronectin does not seem to play a major role in cholesterol gallstone disease.

Impaired human gallbladder lipid absorption in cholesterol gallstone disease and its effect on cholesterol solubility in bile

2000 ◽  
Vol 118 (5) ◽  
pp. 912-920 ◽  
Author(s):  
Stefano Ginanni Corradini ◽  
Walter Elisei ◽  
Luca Giovannelli ◽  
Cristina Ripani ◽  
Paola Della Guardia ◽  
...  
Keyword(s):  
Cholesterol Gallstone ◽  
Gallstone Disease ◽  
Lipid Absorption ◽  
Human Gallbladder ◽  
Cholesterol Solubility

Zebrafish tyrosylprotein sulfotransferase: molecular cloning, expression, and functional characterization

2004 ◽  
Vol 82 (2) ◽  
pp. 295-303 ◽  
Author(s):  
Emi Mishiro ◽  
Ming-Yih Liu ◽  
Yoichi Sakakibara ◽  
Masahito Suiko ◽  
Ming-Cheh Liu
Keyword(s):  
Molecular Cloning ◽  
Divalent Cations ◽  
Functional Characterization ◽  
The Other ◽  
Inhibitory Effects ◽  
Amino Acid Sequence Level ◽  
Ph Optimum ◽  
Rapid Amplification ◽  
High Degree

By employing the reverse transcriptase – polymerase chain reaction technique in conjunction with 3' rapid amplification of cDNA ends, a full-length cDNA encoding a zebrafish (Danio rerio) tyrosylprotein sulfotransferase (TPST) was cloned and sequenced. Sequence analysis revealed that this zebrafish TPST is, at the amino acid sequence level, 66% and 60% identical to the human and mouse TPST-1 and TPST-2, respectively. The recombinant form of the zebrafish TPST, expressed in COS-7 cells, exhibited a pH optimum at 5.75. Manganese appeared to exert a stimulatory effect on the zebrafish TPST. The activity of the enzyme determined in the presence of 20 mM MnCl2 was more than 2.5 times that determined in the absence of MnCl2. Of the other nine divalent metal cations tested at a 10 mM concentration, Co2+ also showed a considerable stimulatory effect, while Ca2+, Pb2+, and Cd2+ exerted some inhibitory effects. The other four divalent cations, Fe2+, Cu2+, Zn2+, and Hg2+, inhibited completely the sulfating activity of the zebrafish TPST. Using the wild-type and mutated P-selectin glycoprotein ligand-1 N-terminal peptides as substrates, the zebrafish TPST was shown to exhibit a high degree of substrate specificity for the tyrosine residue on the C-terminal side of the peptide. These results constitute a first study on the cloning, expression, and characterization of a zebrafish cytosolic TPST.Key words: zebra fish, tyrosylprotein sulfotransferase, molecular cloning.

scholarly journals Cloning, expression, and characterization of the human eosinophil eotaxin receptor.

1996 ◽  
Vol 183 (5) ◽  
pp. 2349-2354 ◽  
Author(s):  
B L Daugherty ◽  
S J Siciliano ◽  
J A DeMartino ◽  
L Malkowitz ◽  
A Sirotina ◽  
...  
Keyword(s):  
Chemokine Receptor ◽  
Allergic Inflammation ◽  
The Other ◽  
Binding Affinities ◽  
Binding Studies ◽  
Human Eosinophil ◽  
Lower Affinity ◽  
Competition Binding ◽  
G Protein Coupled

Although there is a mounting body of evidence that eosinophils are recruited to sites of allergic inflammation by a number of beta-chemokines, particularly eotaxin and RANTES, the receptor that mediates these actions has not been identified. We have now cloned a G protein-coupled receptor, CC CKR3, from human eosinophils which, when stably expressed in AML14.3D10 cells bound eotaxin, MCP-3 and RANTES with Kds of 0.1, 2.7 and 3.1 nM, respectively. CC CKR3 also bound MCP-1 with lower affinity, but did not bind MIP-1 alpha or MIP-1 beta. Eotaxin, RANTES, and to a lessor extent MCP-3, but not the other chemokines, activated CC CKR3 as determined by their ability to stimulate a Ca(2+) -flux. Competition binding studies on primary eosinophils gave binding affinities for the different chemokines which were indistinguishable from those measured with CC CKR3. Since CC CKR3 is prominently expressed in eosinophils we conclude that CC CKR3 is the eosinophil eotaxin receptor. Eosinophils also express a much lower level of a second chemokine receptor, CC CKR1, which appears to be responsible for the effects of MIP-1 alpha.

scholarly journals Resolution and functional characterization of two mitochondrial iron-sulphur centres of the ‘high-potential iron-sulphur protein’ type

1976 ◽  
Vol 153 (1) ◽  
pp. 39-48 ◽  
Author(s):  
T Ohnishi ◽  
W J Ingledew ◽  
S Shiraishi
Keyword(s):  
Line Shape ◽  
Mitochondrial Membrane ◽  
Physiological Function ◽  
Functional Characterization ◽  
High Potential ◽  
The Other ◽  
Power Dependence ◽  
Iron Protein ◽  
Kinetic Behaviour

Two distinct iron-sulphur centres of the ‘HiPIP’ (high-potential iron-protein) type are distinguished in both pigeon heart and ox heart mitochondria. These two species, although both are paramagnetic in the oxidized state, exhibit signals which differ in their detailed line shape, field position, and temperature- and power-dependence. They also exhibit different thermodynamic and kinetic behaviour and are located on opposite sides of the mitochondrial coupling membrane. One of these centres corresponds to Centre S-3. The other ‘HiPIP’-type centre is removed readily from the mitochondrial membrane and its physiological function is not known.

scholarly journals Heterogeneous clinical features in Cockayne syndrome-A patients with the same mutation and in siblings

2021 ◽  
Author(s):  
Asma CHIKHAOUI ◽  
Ichraf Kraoua ◽  
Nadège Calmels ◽  
Sami Bouchoucha ◽  
Cathy Obringer ◽  
...  
Keyword(s):  
North Africa ◽  
Clinical Symptoms ◽  
Excision Repair ◽  
Functional Characterization ◽  
Clinical Manifestations ◽  
Case Series ◽  
Cockayne Syndrome ◽  
The Other ◽  
Indel Mutation

Abstract Background Cockayne syndrome (CS) is a rare autosomal recessive disorder caused by mutations in ERCC6/CSB or ERCC8/CSA that participate in transcription-coupled nucleotide excision repair (TC-NER) of UV-induced DNA damage. CS patients display a large heterogeneity of clinical symptoms and severities, the reason of which is not fully understood, and little data is available for affected siblings. CS is largely undiagnosed in North Africa. Methods We report here the clinical description as well as genetic and functional characterization of eight North African CS patients, including siblings. These patients, who belonged to six unrelated families, underwent complete clinical examination and biochemical analyses. Sanger sequencing was performed for the recurrent mutation in five families, and targeted gene sequencing for one patient of the other family. We also performed RRS (Recovery RNA Synthesis) to confirm the functional impairment of DNA repair in the identified mutations. Results Six out of eight patients carried a homozygous indel mutation (c.598_600delinsAA) in exon 7 of ERCC8, and displayed a variable clinical spectrum, including between siblings, despite sharing the same mutation. The other two patients were Tunisian siblings who carried a homozygous splice-site variant in ERCC8 (c.843 + 1 G > C). They presented more severe clinical manifestations, which are in general rarely associated with CSA mutations, leading to gastrostomy and hepatic damage. Impaired TC-NER was confirmed by RRS in six tested patients. Conclusions This study provides the first deep characterization of case series of rare CS-A patients in North Africa. They carry mutations described to date only in this region and the Middle-East. We also provide the largest characterization of unrelated patients, as well as siblings, with the same mutation, providing a framework for dissecting elusive genotype-phenotype correlations in CS.

scholarly journals Green fluorescent protein-based assays for high-throughput functional characterization and ligand-binding studies of biotin protein ligase

2016 ◽  
Vol 8 (2) ◽  
pp. 418-424 ◽  
Author(s):  
Samuel P. Askin ◽  
Thomas E. H. Bond ◽  
Patrick M. Schaeffer
Keyword(s):  
Green Fluorescent Protein ◽  
Ligand Binding ◽  
High Throughput ◽  
Fluorescent Protein ◽  
Functional Characterization ◽  
Binding Studies ◽  
Biotin Protein Ligase ◽  
Green Fluorescent

Rapid functional characterization of GFP-tagged biotin protein ligase (BirA-GFP) with a high-throughput DSF-GTP assay.

scholarly journals Isolation and chemical and immunochemical characterization of the peanut-lectin-binding glycoprotein from human milk-fat-globule membranes

1984 ◽  
Vol 224 (2) ◽  
pp. 581-589 ◽  
Author(s):  
J Fischer ◽  
P J Klein ◽  
G H Farrar ◽  
F G Hanisch ◽  
G Uhlenbruck
Keyword(s):  
Human Milk ◽  
Binding Sites ◽  
Milk Fat ◽  
Lectin Binding ◽  
Functional Characterization ◽  
Milk Fat Globule ◽  
Fat Globule ◽  
Milk Fat Globule Membranes ◽  
Human Milk Fat

Membrane glycoprotein with high Mr (HMr-MGP) was purified from neuraminidase-treated Triton X-100-solubilized human milk-fat-globule membranes by peanut-agglutinin (PNA) affinity chromatography. The high carbohydrate content (75%), blood-group-A activity and typical monosaccharide composition (L-fucose, D-galactose, N-acetyl-D-glucosamine and N-acetyl-D-galactosamine in the proportions 0.26:1.00:1.85:1.30) indicate that the isolated HMr-MGP is a mucinous substance. Fractionation of the oligosaccharides from alkaline-borohydride-treated HMr-MGP on Bio-Gel P-2 suggest that the PNA-binding sites are located mainly on longer (tetra- to deca-saccharide) alkali-labile bound oligosaccharide chains. Polyclonal antibodies raised against the HMr-MGP showed an antigenic distribution in histological sections that was comparable with the distribution of peroxidase-labelled-PNA-binding sites in both normal and malignant breast tissues. The positive immunohistological staining of some other tissue components with this antibody indicates that HMr-MGP is not strictly breast-associated. The functional role of HMr-MGP is unknown, but, since its expression is dependent on the differentiation state of secretory epithelial cells, it serves as a differentiation antigen that can be used for better functional characterization of breast cancers.

scholarly journals FUNCTIONAL PROPERTIES OF TABLE SUGARS DERIVED FROM THE SAP OF THE INFLORESCENCES OF 03 COCONUT (COCOS NUCIFERA.L) CULTIVARS IN COTE DIVOIRE

2020 ◽  
Vol 8 (11) ◽  
pp. 254-263
Author(s):  
Okoma D. Muriel J. ◽  
◽  
Konan K. Jean Louis ◽  
Assa Rebecca R ◽  
◽  
...  
Keyword(s):  
Functional Characterization ◽  
Cane Sugar ◽  
Polyphenolic Compounds ◽  
The Other ◽  
Total Polyphenols ◽  
Coconut Tree ◽  
White Cane ◽  
Crystalline Sugars ◽  
Cooking Times

This study is part of a context of diversification of the uses of the Ivorian coconut tree. The objective was to determine the functional characteristics of crystalline sugars derived from the sap of inflorescences of three coconut cultivars. Red and white sugars from cane were taken as controls. Variations in final temperatures coupled with distinct cooking times were carried out in order to evaluate the effect of the time/temperature couple on the studied parameters. Thus, three different treatments were applied.The functional characterization of the sugars studied shows that coconut sugars are an important source of total polyphenols with levels ranging from 34.64 to 143.12 mg/100g.No polyphenolic compounds were assayed in white cane sugar. Coconut sugars from treatment 1 are less energetic than those from the other two treatments. On the other hand, brown and white sugars from sugar cane are more energetic than those from coconut trees.In view of all the above, coconut sugars, especially those from treatment 1, are natural sweeteners with a low energy value. In addition, they are rich in polyphenols and flavonoids, unlike refined cane sugar and its red counterpart which contains very few nutrients.Thus, coconut sugars produced in Cote dIvoire can be considered as a phytonutrient substitute, capable of replacing sugarcane sugars.

Sign in / Sign up

Export Citation Format

Share Document