A 40-kDa polypeptide from papain digestion of the rabbit intestinal Na+/phosphate cotransporter retains Na+ and phosphate cotransport

2002 ◽  
Vol 401 (1) ◽  
pp. 1-10 ◽  
Author(s):  
Brian E Peerce
Keyword(s):  
Phosphate Cotransport ◽  
Papain Digestion

scholarly journals Metabolism of rat bone proteoglycans in vivo

1983 ◽  
Vol 216 (3) ◽  
pp. 589-596 ◽  
Author(s):  
C W Prince ◽  
F Rahemtulla ◽  
W T Butler
Keyword(s):  
Extraction Procedure ◽  
Total Radioactivity ◽  
Mineralized Tissue ◽  
Guanidinium Chloride ◽  
Disaccharide Composition ◽  
A Minor ◽  
Papain Digestion ◽  
Rat Bone

Former evaluations of the role of proteoglycans in mineralization have neglected to address the possibility that the metabolism of proteoglycans may be of significance in this regard. This problem was studied by using radiolabeling in vivo of rat calvaria with [35Sulphate for 2-72 h and a sequential extraction procedure to yield two pools of newly synthesized proteoglycans: one obtained from non-mineralized tissue by extraction with guanidinium chloride (GdmCl) and another obtained only after demineralization with EDTA. Total radioactivity in calvaria was maximal after 12 h of incorporation, but by 36 h had declined to a level that was about 55-65% of maximum. Radioactivity in the GdmCl extract declined steadily after 12 h, whereas that in the EDTA extract remained constant until 36 h, when it began to increase. Each extract contained a minor proteoglycan that eluted at the void volume (Vo) of a Sepharose CL-6B column. Unlike in the EDTA extract, this proteoglycan gradually disappeared from the GdmCl extract. Each extract also contained a major, smaller proteoglycan, with a Kav. of 0.24 and 0.36 in the GdmCl and EDTA extracts respectively. Papain digestion of each extract yielded glycosaminoglycan chains with Kav. values of 0.32 and 0.50 on CL-6B in the GdmCl and EDTA extracts respectively. Digestion of each extract with chondroitinase ABC and chondroitinase AC showed that the glycosaminoglycans were of similar disaccharide composition, with about 85% being 4-sulphated and the remainder 6-sulphated and/or iduronic acid-containing. These data suggest that about 45% of the newly synthesized proteoglycans are removed from the tissue during the course of mineralization.

Characterization of the effector mechanisms of a transmission-blocking antibody upon differentiation ofPlasmodium bergheigametocytes into ookinetesin vitro

Parasitology ◽  
1994 ◽  
Vol 109 (1) ◽  
pp. 11-17 ◽  
Author(s):  
G. R. R. Ranawaka ◽  
A. R. Alejo-Blanco ◽  
R. E. Sinden
Keyword(s):  
Mechanisms Of Action ◽  
Antibody Activity ◽  
Antibody Dependent Cellular Cytotoxicity ◽  
Maximal Inhibition ◽  
Blocking Antibody ◽  
Transmission Blocking ◽  
Ammonium Sulphate Precipitation ◽  
Papain Digestion ◽  
Peripheral Blood Leucocytes

SUMMARYThe transmission-blocking monoclonal antibody 13.1, which recognizes the ookinete surface antigen Pbs21 ofPlasmodium berghei, and an IgG2a isotype control antibody 26.37 were purified by caprylic acid and ammonium sulphate precipitation. Fab fragments were prepared by papain digestion. IgG but not Fab from antibody 13.1 reduced ookinete formation byP. bergheiin culture by as much as 94% at a concentration of 100 μg/ml. There was little difference in antibody efficacy in the range 6·25–400 μg/ml in this assay. The parasite was most sensitive to antibody activity in the first 6–9 h of culture, i.e. the gamete/zygote and early retort stages. Peripheral blood leucocytes (PBL) were essential to achieve maximal inhibition by mAb 13.1 (activity was abrogated totally if PBL were removed). Together the data suggest that one of the mechanisms of action of this antibody is antibody-mediated PBL killing. Phagocytosis of parasites was noted in these experiments in all cultures. We have not attempted in this study to distinguish between Fc-mediated opsonization, as opposed to antibody-dependent cellular cytotoxicity.

scholarly journals Signaling pathways utilized by PTH and dopamine to inhibit phosphate transport in mouse renal proximal tubule cells

2009 ◽  
Vol 296 (2) ◽  
pp. F355-F361 ◽  
Author(s):  
Rochelle Cunningham ◽  
Rajatsubhra Biswas ◽  
Marc Brazie ◽  
Deborah Steplock ◽  
Shirish Shenolikar ◽  
...  
Keyword(s):  
Proximal Tubule ◽  
Signaling Pathways ◽  
Phosphate Transport ◽  
Inhibitory Effect ◽  
Renal Proximal Tubule ◽  
Proximal Tubule Cells ◽  
Phosphate Cotransport ◽  
Tubule Cells ◽  
Renal Proximal Tubule Cells ◽  
Sodium Phosphate Cotransport

The present experiments were designed to detail factors regulating phosphate transport in cultured mouse proximal tubule cells by determining the response to parathyroid hormone (PTH), dopamine, and second messenger agonists and inhibitors. Both PTH and dopamine inhibited phosphate transport by over 30%. The inhibitory effect of PTH was completely abolished in the presence of chelerythrine, a PKC inhibitor, but not by Rp-cAMP, a PKA inhibitor. By contrast, both chelerythrine and Rp-cAMP blocked the inhibitory effect of dopamine. Chelerythrine inhibited PTH-mediated cAMP accumulation but also blocked the inhibitory effect of 8-bromo-cAMP on phosphate transport. On the other hand, Rp-cAMP had no effect on the ability of DOG, a PKC activator, to inhibit phosphate transport. PD98059, an inhibitor of MAPK, had no effect on PTH- or dopamine-mediated inhibition of sodium-phosphate cotransport. Finally, compared with 8-bromo-cAMP, 8-pCPT-2′- O-Me-cAMP, an activator of EPAC, had no effect on phosphate transport. These results outline significant differences in the signaling pathways utilized by PTH and dopamine to inhibit renal phosphate transport. Our results also suggest that activation of MAPK is not critically involved in PTH- or dopamine-mediated inhibition of phosphate transport in mouse renal proximal tubule cells in culture.

Direct production of Fv-fragments from a family of monoclonal IgGs papain digestion

1991 ◽  
Vol 28 (4-5) ◽  
pp. 383-391 ◽  
Author(s):  
Margaret Ornatowska ◽  
Jay A. Glasel
Keyword(s):  
Direct Production ◽  
Papain Digestion

Ruthenium-Red-Induced Cell Agglutination and Surface Glycoprotein and Muco-Polysaccharide

1973 ◽  
Vol 13 (3) ◽  
pp. 901-911
Author(s):  
KOZO UTSUMI ◽  
TAKUZO ODA
Keyword(s):  
Sialic Acid ◽  
Heparan Sulphate ◽  
Ruthenium Red ◽  
The Other ◽  
Surface Glycoprotein ◽  
Ascites Hepatoma ◽  
Cell Agglutination ◽  
Acidic Polysaccharides ◽  
Binding Substance ◽  
Papain Digestion

Ruthenium red induces the agglutination of ascites hepatoma cells. The agglutination was inhibited by acidic polysaccharides but not by haptenic inhibitors of some plant agglutinins. The agglutination was also abolished by treatment of the cells with a low concentration of papain. Papain digestion causes a decrease in the number of microvilli on the cell surface and the release of carbohydrate-containing red-binding substance. One of them is a heparan sulphate-like mucopolysaccharide and the other is a glycoprotein which contains sialic acid. The ruthenium red-binding fractions inhibit the cell agglutination induced by ruthenium red.

scholarly journals Proteolytic interconversion and N-terminal sequences of the Citrobacter diversus major β-lactamases

1991 ◽  
Vol 275 (3) ◽  
pp. 629-633 ◽  
Author(s):  
N Franceschini ◽  
G Amicosante ◽  
M Perilli ◽  
M Maccarrone ◽  
A Oratore ◽  
...  
Keyword(s):  
Klebsiella Oxytoca ◽  
Limited Proteolysis ◽  
Major Product ◽  
Beta Lactamase ◽  
Amino Acid Residues ◽  
Beta Lactamases ◽  
Citrobacter Diversus ◽  
High Degree ◽  
Papain Digestion ◽  
Degree Of Similarity

The N-terminal sequences of the two major beta-lactamases produced by Citrobacter diversus differed only by the absence of the first residue in form II and the loss of five amino acid residues at the C-terminal end. Limited proteolysis of the homogeneous form I protein yielded a variety of enzymatically active products. In the major product obtained after the action of papain, the first three N-terminal residues of form I had been cleaved, whereas at the C-terminal end the treated enzyme lacked five residues. However, this cannot explain the different behaviours of form I, form II and papain digestion product upon chromatofocusing. Form I, which was sequenced up to position 56, exhibited a very high degree of similarity with a Klebsiella oxytoca beta-lactamase. The determined sequence, which contained the active serine residue, demonstrated that the chromosome-encoded beta-lactamase of Citrobacter diversus belong to class A.

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