Denaturing and non-denaturing microsolution isoelectric focussing to mine the metalloproteome

Metallomics ◽  
2009 ◽  
Vol 1 (6) ◽  
pp. 501 ◽  
Author(s):  
Barbara Pioselli ◽  
Caroline Munro ◽  
Andrea Raab ◽  
Christian L. Deitrich ◽  
Kriangsak Songsrirote ◽  
...  
Keyword(s):  
Isoelectric Focussing

Fibrinogen Bern I: A Hereditary Fibrinogen Variant With Defective Conformational Stabilization By Calcium Ions

1981 ◽  
Author(s):  
C Rupp ◽  
C Kuyas ◽  
A Häberli ◽  
M Furlan ◽  
E A Beck
Keyword(s):  
Calcium Ions ◽  
Gel Electrophoresis ◽  
Calcium Binding ◽  
Negative Charge ◽  
Fibrinopeptide A ◽  
Isoelectric Focussing ◽  
High Calcium ◽  
Two Generations ◽  
Polypeptide Chains ◽  
Conformational Stabilization

Inherited hypodysfibrinogenemia (fibrinogen Bern I) was found in four members (two generations) of a family with no haemorrhagic or thrombotic history. Fibrin aggregation curves (350 nm, 37°C) with patient plasma or purified fibrinogen Bern I, after addition of thrombin, were normal at high calcium concentrations (5mM) but delayed at lower calcium concentrations (≤0.lmM). The release of fibrinopeptide A was normal. Whereas the polypeptide chains of fibrinogen Bern I were indistinguishable from normal fibrinogen by SDS-gel-electrophoresis, an abnormal γ-chain with a decreased negative charge was found by isoelectric focussing.Plasmic degradation o| normal fibrinogen, in the presence of calcium (≥ImM), results in only one terminal D fragment which is stabilized by calcium against further degradation of γ-chains. In contrast, degradation of fibrinogen Bern I, under the same conditions, yielded at least two additional smaller D fragments. In conclusion, fibrinogen Bern I is characterized by defective calcium binding in the D domain of the γ-chain.

Status methodology in isoelectric focussing

1983 ◽  
Vol 2 (9) ◽  
pp. 193-196 ◽  
Author(s):  
Pier Giorgio Righetti
Keyword(s):  
Isoelectric Focussing

Photoactivated linkage of catechol O-methyltransferase and S-adenosyl-L-methionine

1984 ◽  
Vol 62 (10) ◽  
pp. 964-969 ◽  
Author(s):  
Peter H. Yu
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Ultraviolet Light ◽  
Paper Chromatography ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Polyacrylamide Gel ◽  
Comt Inhibitors ◽  
Isoelectric Focussing

The formation of a stably linked complex of tritiated S-adenosyl-L-methionine (AdoMet) and catechol O-methyltransferase (COMT) has been achieved by irradiating the enzyme and ligand in Tris–HCl buffer (pH 7.5) with ultraviolet light at 254 nm. The reaction is specific as shown by a number of criteria. COMT inhibitors such as S-adenosylhomocysteine can block this photoactivated linkage. The [3H]AdoMet–COMT adduct has been shown to be a homogeneous protein by Sephadex gel filtration, sodium dodecyl sulfate – polyacrylamide gel electrophoresis, and isoelectric focussing. After extensive proteolysis of the [3H]AdoMet–COMT adduct with pronase P, one major labelled product was released. This fragment could be separated by paper chromatography and was shown to be chromatographically identical to that released from the [3H]AdoMet – phenylethanolamine N-methyltransferase adduct.

scholarly journals Factor VIII-Like Activity of Basic Amphoteric Polyelectrolytes

1977 ◽  
Author(s):  
T. Exner ◽  
K. A. Rickard ◽  
H. Kronenberg
Keyword(s):  
Factor Viii ◽  
Propionic Acid ◽  
Partial Thromboplastin Time ◽  
Factor Ix ◽  
Contact Activation ◽  
Isoelectric Focussing ◽  
Factor Viii Deficiency ◽  
Condensation Products ◽  
Isoelectric Points ◽  
Carrier Ampholytes

The broad-range Ampholine (LKB) 3.5–10 containing a mixture of materials with isoelectric points between pH 3.5 and 10 was found to correct the prolonged PTTK of factor VIII-deficient plasma, and this effect was investigated. Ampholines which are used as carrier ampholytes or pH gradient stabilizers in isoelectric focussing are condensation products of propionic acid and polyethylene polyamines.The most effective of the commercial Ampholine mixtures were found to be those with the most alkaline isoelectric points. Ampholine 9–11 at 0.2% concentration was found to have factor VIII and factor IX activity equal to that of normal plasma as well as somewhat less activity as factor XI. It had no significant correcting effect on plasmas deficient in factors V, VII, X, II or XII. In contrast to polyglutamic and polyaspartic acids which have also been reported to correct for factor VIII deficiency in the PTTK, Ampholine 9–11 was effective whether added before or after kaolin contact activation of the test plasma. However, Ampholine 9–11 inhibited contact activation of plasma in the absence of kaolin and showed less correcting activity in partial thromboplastin time tests utilizing activators other than kaolin.

scholarly journals Analysis of the Iron-Binding Sites of Transferrin by Isoelectric Focussing

1978 ◽  
Vol 16 (10) ◽  
Author(s):  
H. G. van Eijk ◽  
W. L. van Noort ◽  
M. J. Kroos ◽  
C. van der Heul
Keyword(s):  
Binding Sites ◽  
Iron Binding ◽  
Isoelectric Focussing

Isoelectric focussing of nanogram quantities of ferritin

Pathology ◽  
1982 ◽  
Vol 14 (2) ◽  
pp. 228
Author(s):  
R.W. Blunden ◽  
R.J. Kimber
Keyword(s):  
Isoelectric Focussing
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