Radioimmunoassay Studies of Intestinal Calcium-binding Protein in the Pig. II. The Distribution of Intestinal CaBP in Pig Tissues

1975 ◽  
Vol 53 (6) ◽  
pp. 1135-1140 ◽  
Author(s):  
B. M. Arnold ◽  
M. Kuttner ◽  
D. M. Willis ◽  
A. J. W. Hitchman ◽  
J. E. Harrison ◽  
...  
Keyword(s):  
Small Intestine ◽  
Calcium Binding ◽  
Binding Protein ◽  
Gel Filtration ◽  
Protein S ◽  
Binding Activity ◽  
Calcium Binding Protein ◽  
Distal Small Intestine ◽  
Specific Radioimmunoassay ◽  
Kidney Liver

Using a specific radioimmunoassay for porcine intestinal calcium-binding protein (CaBP), we have measured the concentration of CaBP in the various tissues and organs of normal pigs. Intestinal CaBP was present in highest concentration in the upper small intestine, with lower concentrations in the distal small intestine. Intestinal CaBP was also found, in lower concentrations, in kidney, liver, thyroid, pancreas, and blood. In all other tissues, including parathyroid, bone, skeletal muscle, and brain, CaBP immunoreactivity was undetectable or less than in blood. The elution profile of calcium-binding activity and immunoreactivity from gel filtration analysis of kidney and parathyroid extracts suggest that the calcium-binding protein in the parathyroid gland, and the major calcium-binding protein(s) in the kidney, are chemically and immunochemically different from intestinal CaBP.

scholarly journals Nonstructural 5A Protein of Hepatitis C Virus Regulates Soluble Resistance-Related Calcium-Binding Protein Activity for Viral Propagation

2015 ◽  
Vol 90 (6) ◽  
pp. 2794-2805 ◽  
Author(s):  
Giao V. Q. Tran ◽  
Trang T. D. Luong ◽  
Eun-Mee Park ◽  
Jong-Wook Kim ◽  
Jae-Woong Choi ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Protein Interaction ◽  
Calcium Binding ◽  
Crucial Role ◽  
Binding Protein ◽  
Binding Activity ◽  
Calcium Binding Protein ◽  
Virus Propagation ◽  
Ns5a Protein

ABSTRACTHepatitis C virus (HCV) is a major cause of chronic liver disease and is highly dependent on cellular proteins for virus propagation. To identify the cellular factors involved in HCV propagation, we recently performed protein microarray assays using the HCV nonstructural 5A (NS5A) protein as a probe. Of 90 cellular protein candidates, we selected the soluble resistance-related calcium-binding protein (sorcin) for further characterization. Sorcin is a calcium-binding protein and is highly expressed in certain cancer cells. We verified that NS5A interacted with sorcin through domain I of NS5A, and phosphorylation of the threonine residue 155 of sorcin played a crucial role in protein interaction. Small interfering RNA (siRNA)-mediated knockdown of sorcin impaired HCV propagation. Silencing of sorcin expression resulted in a decrease of HCV assembly without affecting HCV RNA and protein levels. We further demonstrated that polo-like kinase 1 (PLK1)-mediated phosphorylation of sorcin was increased by NS5A. We showed that both phosphorylation and calcium-binding activity of sorcin were required for HCV propagation. These data indicate that HCV modulates sorcin activity via NS5A protein for its own propagation.IMPORTANCESorcin is a calcium-binding protein and regulates intracellular calcium homeostasis. HCV NS5A interacts with sorcin, and phosphorylation of sorcin is required for protein interaction. Gene silencing of sorcin impaired HCV propagation at the assembly step of the HCV life cycle. Sorcin is phosphorylated by PLK1 via protein interaction. We showed that sorcin interacted with both NS5A and PLK1, and PLK1-mediated phosphorylation of sorcin was increased by NS5A. Moreover, calcium-binding activity of sorcin played a crucial role in HCV propagation. These data provide evidence that HCV regulates host calcium metabolism for virus propagation, and thus manipulation of sorcin activity may represent a novel therapeutic target for HCV.

scholarly journals The Heparin-Binding Activity of Secreted Modular Calcium-Binding Protein 1 (SMOC-1) Modulates Its Cell Adhesion Properties

PLoS ONE ◽  
2013 ◽  
Vol 8 (2) ◽  
pp. e56839 ◽  
Author(s):  
Marina Klemenčič ◽  
Marko Novinec ◽  
Silke Maier ◽  
Ursula Hartmann ◽  
Brigita Lenarčič
Keyword(s):  
Cell Adhesion ◽  
Calcium Binding ◽  
Binding Protein ◽  
Binding Activity ◽  
Calcium Binding Protein ◽  
Heparin Binding ◽  
Adhesion Properties

scholarly journals Isolation and characterization of grancalcin, a novel 28 kDa EF-hand calcium-binding protein from human neutrophils

1992 ◽  
Vol 286 (2) ◽  
pp. 549-554 ◽  
Author(s):  
C G Teahan ◽  
N F Totty ◽  
A W Segal
Keyword(s):  
Sequence Analysis ◽  
Calcium Binding ◽  
Binding Protein ◽  
Sequence Similarity ◽  
Gel Filtration ◽  
Calcium Binding Protein ◽  
Peptide Sequence ◽  
Human Neutrophils ◽  
Isolation And Characterization ◽  
Ef Hand

A novel 28 kDa protein, which we have named ‘grancalcin’, has been identified in human neutrophils. The protein was isolated from the cytosol and found to be a homodimer, with an apparent molecular mass of 55 kDa on gel filtration. Polyclonal antibodies were raised to the native protein. N-Terminal sequence analysis and tryptic-peptide sequence analysis was performed. The protein exhibits sequence similarity to sorcin, a 24 kDa calcium-binding protein over-expressed in certain multi-drug-resistant cell lines. It appears to be a member of the EF-hand family of calcium-binding proteins. The association of a high proportion of this protein with the membranes and granules in the presence of physiological concentrations of calcium may indicate a role in granule-membrane fusion and degranulation.

scholarly journals The response of the small intestine to vitamin D. Correlation between calcium-binding-protein production and increased calcium absorption

1974 ◽  
Vol 144 (2) ◽  
pp. 339-346 ◽  
Author(s):  
J S Emtage ◽  
D E M Lawson ◽  
E Kodicek
Keyword(s):  
Vitamin D ◽  
Protein Synthesis ◽  
Small Intestine ◽  
Calcium Binding ◽  
Protein Production ◽  
Calcium Absorption ◽  
Binding Protein ◽  
Calcium Binding Protein ◽  
Intestinal Cell ◽  
Stimulation Of

1. The synthesis of calcium-binding protein, a protein produced in the small intestine in response to vitamin D, was investigated with a view to determining whether calcium-binding-protein production could be correlated with the stimulation of calcium absorption by vitamin D. 2. A radioimmunological assay, which can quantitatively estimate calcium-binding-protein concentrations as low as 1μg/g wet wt., was used to detect the synthesis of soluble calcium-binding protein. 3. When used on intestinal supernatants from chicks dosed with vitamin D, calcium-binding protein was not detectable at 8h but was present after 12h at a concentration of 8.6μg/g wet wt.; in agreement with this an increase in calcium absorption due to vitamin D was detected at 12h but not at 8h. 4. The synthesis of calcium-binding protein was also monitored directly by making use of the ability of the iodinated antiserum to bind specifically to nascent calcium-binding protein chains on intestinal polyribosomes; in this way calcium-binding-protein synthesis could be detected 8h after dosage with vitamin D. Further, the binding reaction indicated a near linear increase in the calcium-binding-protein-synthesizing capacity over a 16h period. 5. From the amount of calcium-binding protein present 12 and 24h after vitamin D administration it is calculated that calcium-binding-protein mRNA is produced at approx. 1mol/min per intestinal cell. 6. It is concluded that the high correlation between the initiation of calcium-binding-protein synthesis and the stimulation of calcium absorption by vitamin D strengthens the proposal that calcium-binding protein plays an important role in calcium transport.

Radioimmunoassay Studies of Intestinal Calcium-binding Protein in the Pig. I. Identification of Intestinal Calcium-binding Protein in Blood and Response to a Low Calcium Diet

1975 ◽  
Vol 53 (6) ◽  
pp. 1129-1134 ◽  
Author(s):  
B. M. Arnold ◽  
M. Kuttner ◽  
R. Swaminathan ◽  
A. D. Care ◽  
A. J. W. Hitchman ◽  
...  
Keyword(s):  
Small Intestine ◽  
Intestinal Mucosa ◽  
Calcium Binding ◽  
Binding Protein ◽  
Calcium Binding Protein ◽  
Intestinal Biopsy ◽  
Mixed Venous Blood ◽  
Calcium Diet ◽  
Low Calcium Diet ◽  
Low Calcium

We have developed a radioimmunoassay for porcine intestinal calcium-binding protein (CaBP) and have used it to detect CaBP in pig plasma. Plasma CaBP is identical to intestinal CaBP on the basis of immunological activity, molecular size, and molecular charge properties. The plasma CaBP concentration was greater in the portal blood than in mixed venous blood, suggesting that blood CaBP originates in the gut. Two of four 15-week-old littermate pigs were placed on a low calcium diet (0.15% calcium, 0.65% phosphorus) and two on a control diet (0.65% calcium, 0.65% phosphorus). After 2 weeks, the entire small intestine was removed and divided into nine 1.8-m segments. CaBP was assayed in both plasma and intestinal mucosa. When the two pigs on a low calcium diet were compared with two control pigs, there was a general increase in immunoreactive CaBP in both plasma and intestinal mucosa. However, there was no increment in immunoreactive CaBP in the first 1.8-m segment of small intestine. Seventy-one percent of the increment in CaBP occurred distal to the first two segments. The largest fractional low calcium diet effect occurred in the ileum. The mean CaBP concentration for the total small intestine increased by a factor of 1.9. The plasma CaBP concentration increased by a factor of 2.6. In these pigs, plasma CaBP was a more reliable indicator of change in CaBP status than was the measurement in the proximal gut segment which contained the duodenum. The assay of CaBP in blood is convenient and may obviate the sampling errors inherent in intestinal biopsy.

scholarly journals Identification and characterization of a calcium-binding protein in the mouse chorioallantoic placenta

1986 ◽  
Vol 233 (1) ◽  
pp. 41-49 ◽  
Author(s):  
R S Tuan ◽  
S T Cavanaugh
Keyword(s):  
Calcium Binding ◽  
Binding Protein ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Calcium Binding Protein ◽  
Chorionic Villi ◽  
Chorioallantoic Placenta ◽  
Identification And Characterization

Mouse chorioallantoic placenta contains a specific calcium-binding protein (MCaBP). A procedure involving gel filtration and ion-exchange chromatography was developed to purify the MCaBP. The MCaBP activity increased as a function of embryonic gestation and was highly specific for Ca2+. The MCaBP is a monomeric protein of Mr 57000, with pI 4.7. Specific antibodies were prepared against the MCaBP and were used to localize the MCaBP to syncytiotrophoblasts of the chorionic villi of mouse chorioallantoic placenta. These properties suggest that the MCaBP may be involved in transplacental calcium transport.

Usefulness of gel filtration fraction as potential biomarker for neurocysticercosis in serum: towards a new diagnostic tool

Parasitology ◽  
2016 ◽  
Vol 144 (4) ◽  
pp. 426-435 ◽  
Author(s):  
D. S. NUNES ◽  
H. T. GONZAGA ◽  
V. S. RIBEIRO ◽  
J. P. CUNHA-JÚNIOR ◽  
J. M. COSTA-CRUZ
Keyword(s):  
B Cell ◽  
Calcium Binding ◽  
Binding Protein ◽  
Gel Filtration ◽  
Calcium Binding Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Filtration Fraction ◽  
B Cell Epitopes ◽  
Diagnostic Parameters

SUMMARYThere is an increasing interest in improving neurocysticercosis (NCC) diagnosis through the search of new and alternative antigenic sources, as those obtained from heterologous antigens. The aim of this study was to obtain potential biomarkers for NCC diagnosis after gel filtration chromatography [gel filtration fraction (GFF)] from the total saline extract (SE) from Taenia saginata metacestodes, followed by protein identification and application in immunodiagnostic. SE and GFF proteic profiles were characterized in gel electrophoresis, and diagnostic performance was verified by testing 160 serum samples through enzyme-linked immunosorbent assay and immunoblotting. Sensitivity (Se), specificity (Sp) and other diagnostic parameters were calculated. Polypeptides of interest in the diagnosis of human NCC present at GFF were analysed by mass spectrometry (MS) and B-cell epitopes were predicted. GFF had the best diagnostic parameters: Se 93·3%; Sp 93%; AUC 0·990; LR+ = 13·42 and LR− = 0·07, and proved to be useful reacting with serum samples in immunoblotting. Proteic profile ranged from 64 to 68 kDa and enolase and calcium binding protein calreticulin precursor were identified after MS. The enolase and calcium-binding protein calreticulin precursor showed 18 and 10 predicted B-cell epitopes, respectively. In conclusion we identified important markers in the GFF with high efficiency to diagnose NCC.

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