scholarly journals Messenger ribonucleic acids from pig intestinal mucosa direct synthesis of calcium-binding protein in a cell-free translation system

1980 ◽  
Vol 185 (3) ◽  
pp. 601-607 ◽  
Author(s):  
H Mellersh ◽  
S Tomlinson ◽  
A Pollock
Keyword(s):  
Calcium Binding ◽  
Binding Protein ◽  
Direct Synthesis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Duodenal Mucosa ◽  
Exchange Chromatography ◽  
Calcium Binding Protein ◽  
Translation System ◽  
Free System

mRNA from pig duodenal mucosa directs synthesis, in a wheat-germ cell-free system, of two products precipitated by antiserum to pure calcium-binding protein. One of these products has a higher molecular weight than authentic calcium-binding protein, but, like the authentic protein, is heat-stable. The other protein co-migrates with pure calcium-binding protein on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, is stable on heating to 70 degrees C for 15 min and alters its elution position on ion-exchange chromatography depending on whether Ca2+ is present in or absent from the elution buffer. The synthesized protein has these properties in common with authentic calcium-binding protein.

scholarly journals Calcium-binding protein in the duodenal mucosa of uremic patients and normal subjects

1975 ◽  
Vol 8 (2) ◽  
pp. 110-118 ◽  
Author(s):  
Peter Piazolo ◽  
Jürgen Hotz ◽  
Klaus Helmke ◽  
Hans-Eduard Franz ◽  
Mark Schleyer
Keyword(s):  
Calcium Binding ◽  
Binding Protein ◽  
Normal Subjects ◽  
Duodenal Mucosa ◽  
Calcium Binding Protein ◽  
Uremic Patients

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.

scholarly journals The interaction of the calcium-binding protein (troponin C) with bivalent cations and the inhibitory protein (troponin I)

1974 ◽  
Vol 137 (2) ◽  
pp. 145-154 ◽  
Author(s):  
J. F. Head ◽  
S. V. Perry
Keyword(s):  
Skeletal Muscle ◽  
Calcium Binding ◽  
Troponin I ◽  
Binding Protein ◽  
Sodium Dodecyl ◽  
Psoas Muscle ◽  
Calcium Binding Protein ◽  
Sedimentation Equilibrium ◽  
Free Form ◽  
Inhibitory Protein

1. The molecular weight of the calcium-binding protein of rabbit white skeletal muscle was estimated to be 18500 by sedimentation equilibrium and electrophoresis in sodium dodecyl sulphate. 2. Addition of 2 Ca2+ ions per molecule produced reversible changes in the u.v.-absorption spectrum that are interpreted as arising from conformational changes in the structure of the protein. 3. Cd2+ was almost as effective as Ca2+ in producing the spectral changes. Other bivalent metal ions, particularly Mg2+, were less effective. 4. Binding of Ca2+ by the calcium-binding protein produced an increase in mobility to the anode on electrophoresis in 6m-urea at pH8.6. The Ca2+-saturated form of the protein was more retarded on gel filtration than the Ca2+-free form. 5. In the presence of Ca2+ the calcium-binding protein formed an equimolar complex with the inhibitory protein. This complex was stable in 8m-urea and in the pH range 7.0–8.6. 6. An isotope-dilution method for the measurement of the content of calcium-binding protein in whole muscle is described. In rabbit psoas muscle the ratio of actin monomers to molecules of calcium-binding protein was approx. 7:1. Similar values were obtained for red skeletal and cardiac muscle. 7. Evidence is presented indicating that in the rabbit the inhibitory protein of the troponin complex of red skeletal and cardiac muscles is different from the inhibitory protein of white skeletal muscle.

scholarly journals Stimulation of intestinal calcium-binding-protein mRNA synthesis in the nucleus of vitamin D-deficient chicks by 1,25-dihydroxycholecalciferol

1978 ◽  
Vol 175 (3) ◽  
pp. 1089-1094 ◽  
Author(s):  
R. Spencer ◽  
M. Charman ◽  
D. E. M. Lawson
Keyword(s):  
Vitamin D ◽  
Calcium Binding ◽  
Binding Protein ◽  
Calcium Binding Protein ◽  
Free System ◽  
Rna Molecules ◽  
Pulse Dose ◽  
A Cell ◽  
Active Calcium ◽  
Stimulation Of

Stimulation of intestinal calcium transport by the hormone 1,25-dihydroxycholecalciferol appears to involve RNA transcriptions and the synthesis of new proteins. Although one of these proteins has been identified as calcium-binding protein, no RNA molecules specifically induced by the hormone in the nucleus have been identified. Nuclear RNA from intestine of vitamin D-deficient chicks before and at various time intervals after treatment with the hormone or cholecalciferol was tested for its ability to code for calcium-binding protein in a cell-free system. Calcium-binding-protein mRNA could only just be detected in the intestinal nuclei 2h after dosing with these steroids which is the same time that it was first observed in the polyribosomes. Thus 1,25-dihydroxycholecalciferol induces the production of new calcium-binding protein by stimulating the formation and rapid release from the nucleus of new mRNA molecules for this protein. Polyribosomal translation of the mRNA continued only as long as it was being synthesized, and the maximum rate of synthesis following a pulse dose of 125ng of the hormone was the same as that observed after prolonged stimulation with cholecalciferol. The possibility that other 1,25-dihydroxycholecalciferol-dependent events may be occurring in the nucleus in the lag period between accumulation of the hormone in the intestine and the appearance of active calcium-binding-protein mRNA, and that these may ultimately control the synthesis of that mRNA, is discussed.

Interaction of the proteasome S5a/Rpn10 multiubiquitin-binding protein and the 8 kDa calcium-binding protein of Schistosoma mansoni

Parasitology ◽  
2003 ◽  
Vol 127 (4) ◽  
pp. 337-347 ◽  
Author(s):  
D. RAM ◽  
E. ZIV ◽  
F. LANTNER ◽  
I. SCHECHTER
Keyword(s):  
Calcium Binding ◽  
Binding Protein ◽  
Calcium Binding Protein ◽  
26S Proteasome ◽  
Dependent Manner ◽  
Terminal Portion ◽  
Binding Interaction ◽  
Western Blots ◽  
Free System ◽  
Amino Terminal

A distinct 8 kDa calcium-binding protein (CaBP) is preferentially expressed at the cercarial stage during the life-cycle of the schistosome. Available data indicate that this CaBP may be associated with tissue/organ remodelling (involving protein degradation and synthesis of new proteins) during transformation of the cercariae from free-living form in water to parasitic life in the vertebrate host. Many CaBP molecules (e.g. calmodulin) show Ca++-dependent interaction with target proteins and thus modulate their activity. Accordingly, the parasite 8 kDa CaBP was used as a probe to clone and identify putative target protein(s) directly by binding interaction. Screening of schistosome λgt11 expression library with radio-iodinated CaBP yielded several overlapping clones showing Ca++-dependent binding of the CaBP. Sequence analyses revealed that these clones encode the S5a/Rpn10 multiubiquitin-binding protein which is a component of the regulatory 19S subunit of the 26S proteasome. The schistosome molecule, designated SmS5a, is 420 amino acids long. The nearly full length molecule (Gln3–Ser420) as well as the amino terminal (N-S5a, Gln3–Gly200) and carboxyl-terminal (C-S5a, Asp225–Ser420) portions were synthesized in bacteria, purified, and antibodies to the parasite SmS5a were prepared. Interaction between SmS5a and the 8 kDa CaBP in a Ca++-dependent manner was found under various experimental conditions: CaBP-Sepharose bound soluble SmS5a, immobilized SmS5a bound soluble CaBP, and complex formation was found when both molecules were in solution. Furthermore, it was shown that the C-terminal portion of SmS5a, but not the N-terminal portion of the molecule, reacted with the CaBP. SmS5a synthesized in a cell-free system and Western blots revealed 2 species, conceivably corresponding to the naked molecule (~50 kDa) and the molecule subjected to post-translational modification (~70 kDa). The present studies suggest that proteasome activity may be modulated by calcium, and this modulation is mediated via CaBP molecule(s).

Presence of two new fatty acid binding proteins in catfish liver

1996 ◽  
Vol 74 (5) ◽  
pp. 675-680 ◽  
Author(s):  
Santiago M. Di Pietro ◽  
José A. Santomé
Keyword(s):  
Fatty Acid ◽  
Fatty Acid Binding Protein ◽  
Binding Protein ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Exchange Chromatography ◽  
Amino Acid Sequences ◽  
Fatty Acid Binding ◽  
Acid Binding Protein ◽  
Bovine Heart

A basic fatty acid binding protein (FABP), closely related to that of chicken liver, was isolated and characterized from catfish (Rhamdia sapo) liver in a previous work. Results herein show the presence of another two FABPs in which partial amino acid sequences reveal great similarity with the corresponding sequences of other already known FABPs belonging to the heart type. The purification procedures for both proteins involve gel filtration, anion-exchange chromatography, and sodium dodecyl sulfate – polyacrylamide gel electrophoresis (as a last step). Because both FABP N-termini were blocked, they were submitted to in-gel tryptic digestion and the resulting peptides were separated by high performance liquid chromatography, and sequenced by Edman degradation. One of these proteins presented the highest identity percentage when compared with those of the human and bovine heart and bovine brain (81%), and the other when compared with those of chicken retina (75%) and mouse and bovine heart FABP (70%). The presence of several FABPs plus the fact that they belong to different types, as found in the Rhamdia sapo liver, is unusual in mammals, which express a characteristic liver-type member of this protein family.Key words: fatty acid binding protein, liver, catfish, Rhamdia sapo.

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