scholarly journals The purification and characterization of bovine C4, the fourth component of complement

1979 ◽  
Vol 177 (3) ◽  
pp. 959-965 ◽  
Author(s):  
N A Booth ◽  
R D Campbell ◽  
J E Fothergill
Keyword(s):  
High Yield ◽  
Haemolytic Activity ◽  
Purification And Characterization ◽  
Bovine Plasma ◽  
Beta Globulin ◽  
Alpha Beta ◽  
Fourth Component ◽  
Polypeptide Chains ◽  
Human Component

The fourth component of complement, C4, was isolated from bovine plasma in high yield, by using simple purification techniques. The protein, like human component C4, is a beta-globulin with a mol.wt. of about 200 000 and consists of three polypeptide chains, alpha, beta and gamma, with apparent mol. wts. of 98 000, 82 000 and 32 000 respectively. The chains of C4 have been separated by methods previously used for human C4. Their amino acid compositions are very similar to those of the human component, but differences in carbohydrate distribution have been observed. The haemolytic activity of bovine C4 is totally destroyed by incubation with bovine C1s, the activated subcomponent of the first component of complement. Component C4, treated in this way, was shown to be cleaved in the alpha chain, which was decreased in mol.wt. by about 9000, corresponding to the removal of subcomponent C4a.

scholarly journals Purification and characterization of subcomponent C1q of the first component of bovine complement

1979 ◽  
Vol 177 (2) ◽  
pp. 531-540 ◽  
Author(s):  
R. Duncan Campbell ◽  
Nuala A. Booth ◽  
John E. Fothergill
Keyword(s):  
Sodium Dodecyl ◽  
Molar Ratio ◽  
High Yield ◽  
Molecular Characteristics ◽  
Haemolytic Activity ◽  
Reducing Conditions ◽  
Equilibrium Sedimentation ◽  
Triple Helices ◽  
Polypeptide Chains

Bovine C1q, a subcomponent of the first component of complement, was purified in high yield by a combination of euglobulin precipitation, and ion-exchange and molecularsieve chromatography on CM-cellulose and Ultrogel AcA 34. Approx. 12–16mg can be isolated from 1 litre of serum, representing a yield of 13–18%. The molecular weight of undissociated subcomponent C1q, as determined by equilibrium sedimentation, is 430000. On sodium dodecyl sulphate/polyacrylamide gels under non-reducing conditions, subcomponent C1q was shown to consist of two subunits of mol.wts. 69000 and 62000 in a molar ratio of 2:1. On reduction, the 69000-mol.wt. subunit gave chains of mol.wts. 30000 and 25000 in equimolar ratio, and the 62000-mol.wt. subunit decreased to 25000. The amino acid composition, with a high value for glycine, and the presence of hydroxyproline and hydroxylysine, suggests that there is a region of collagen-like sequence in the molecule. This is supported by the loss of haemolytic activity and the degradation of the polypeptide chains of subcomponent C1q when digested by collagenase. All of these molecular characteristics support the structure of six subunits, each containing three different polypeptide chains, with globular heads connected by collagen triple helices as proposed by Reid & Porter (1976) (Biochem. J.155, 19–23) for human subcomponent C1q. Subcomponent C1q contains approx. 9% carbohydrate; analysis of the degree of substitution of the hydroxylysine residues revealed that 91% are modified by the addition of the disaccharide unit Gal-Glc. Bovine subcomponent C1q generates full C1 haemolytic activity when assayed with human subcomponents C1r and C1s.

High-yield purification and characterization of recombinant human leukotactin-1 inPichia pastoris

2004 ◽  
Vol 9 (1) ◽  
pp. 1-6 ◽  
Author(s):  
In Hwan Lim ◽  
Kong Ju Lee ◽  
Eun Kyoung Lee ◽  
Mu Rim Choi ◽  
Gue-Wha Lee ◽  
...  
Keyword(s):  
High Yield ◽  
Purification And Characterization

High-Yield Purification and Characterization of Human Asialoglycoprotein Receptor

1995 ◽  
Vol 6 (3) ◽  
pp. 251-255 ◽  
Author(s):  
U. Treichel ◽  
T. Schreiter ◽  
K.H.M. Zumbuschenfelde ◽  
R.J. Stockert
Keyword(s):  
Asialoglycoprotein Receptor ◽  
High Yield ◽  
Purification And Characterization

scholarly journals Simultaneous purification and characterization of glucokinase, fructokinase and glucose-6-phosphate dehydrogenase from Zymomonas mobilis

1985 ◽  
Vol 228 (3) ◽  
pp. 627-634 ◽  
Author(s):  
R K Scopes ◽  
V Testolin ◽  
A Stoter ◽  
K Griffiths-Smith ◽  
E M Algar
Keyword(s):  
Kinetic Parameters ◽  
Zymomonas Mobilis ◽  
High Yield ◽  
Purification And Characterization ◽  
Glucose 6 Phosphate Dehydrogenase

The three enzymes glucokinase (EC 2.7.1.2), fructokinase (EC 2.7.1.4) and glucose-6-phosphate dehydrogenase (EC 1.1.1.49) were isolated in high yield from extracts of Zymomonas mobilis. The principal steps in the isolation procedures involved the use of selected dye-ligand adsorbent columns, with affinity elution of two of the three enzymes. Glucokinase and fructokinase are dimeric proteins (2 × 33000 Da and 2 × 28000 Da respectively) and glucose-6-phosphate dehydrogenase is a tetramer (4 × 52000 Da). Some similarities in the structural and kinetic parameters of the two kinases were noted, but they have absolute specificity for their substrates. Fructokinase is strongly inhibited by glucose; otherwise non-substrate sugars had little effect on any of the three enzymes.

scholarly journals Structural and functional differences between the H-2 controlled Ss and Slp proteins.

1978 ◽  
Vol 148 (5) ◽  
pp. 1186-1197 ◽  
Author(s):  
A Ferreira ◽  
V Nussenzweig ◽  
I Gigli
Keyword(s):  
Hemolytic Activity ◽  
Structural Data ◽  
Serum Levels ◽  
Alpha Chain ◽  
Functional Differences ◽  
Alpha Beta ◽  
Fourth Component ◽  
Covalently Linked ◽  
Polypeptide Chains ◽  
High Degree

Based on functional and structural data, it is concluded that the Ss protein in the mouse expresses the activity of the fourth component of complement. Removal of the Ss, but not of Slp, antigen correlates with a high degree of significance (P less than 0.001) with decrease of C4 hemolytic activity. In phenotypically Slp negative mice the plasma/serum levels of Ss correlate with the C4 activity (P less than 0.001). Structurally, Ss is a 209,000-mol wt protein, consisting of three covalently linked polypeptide chains (alpha,beta,gamma). Treatment of Ss with C1 cleaves a 7,000-8,000-mol wt fragment from the alpha-chain. Slp is also a three chain covalently linked protein of 209,000 daltons, however its three chains differ in size from those of the Ss protein. Slp does not express hemolytic activity and its alpha-chain is not cleaved by C1.

High-yield expression in Escherichia coli, purification, and characterization of properly folded major peanut allergen Ara h 2

2003 ◽  
Vol 31 (2) ◽  
pp. 250-259 ◽  
Author(s):  
Katrin Lehmann ◽  
Silke Hoffmann ◽  
Philipp Neudecker ◽  
Martin Suhr ◽  
Wolf-Meinhard Becker ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
High Yield ◽  
Purification And Characterization ◽  
Peanut Allergen ◽  
Ara H 2 ◽  
Expression In Escherichia Coli

High-yield expression, purification and characterization of tumor-targeted IFN-α2a

Cytotherapy ◽  
2007 ◽  
Vol 9 (1) ◽  
pp. 60-68 ◽  
Author(s):  
J. Meng ◽  
Z. Yan ◽  
J. Wu ◽  
L. Li ◽  
X. Xue ◽  
...  
Keyword(s):  
High Yield ◽  
Purification And Characterization
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