scholarly journals Isolation and characterization of rat plasma fibronectin

1981 ◽  
Vol 197 (3) ◽  
pp. 529-534 ◽  
Author(s):  
R E Weiss ◽  
A H Reddi
Keyword(s):  
Proteinase Inhibitors ◽  
Rat Plasma ◽  
Proteolytic Degradation ◽  
Affinity Column ◽  
Plasma Fibronectin ◽  
Collagen Binding ◽  
Isolation And Characterization ◽  
Monospecific Antibodies ◽  
Shed Light

Rat plasma fibronectin has been isolated and characterized and monospecific antibodies were prepared to it. Two components of fresh rat plasma (in the presence of proteinase inhibitors) bound to a gelatin-Sepharose affinity column. One protein was eluted with 4.0 M-urea and was identified as fibronectin. Another protein was eluted from the gelatin-Sepharose column with 8.0 M-urea and was identified as a 70 000-Mr collagen-binding molecule. This 70 000-Mr fragment was found to be a normal constituent of blood plasma, and its presence did not represent a proteolytic degradation product formed during isolation. The antibodies prepared against rat fibronectin only weakly cross-reacted with plasma fibronectins of chicken, horse and human. These studies shed light on the metabolic interrelationships between fibronectin and other collagen-binding molecules.

scholarly journals Carbohydrate binding activities of Bradyrhizobium japonicum. II. Isolation and characterization of a galactose-specific lectin.

1990 ◽  
Vol 111 (4) ◽  
pp. 1639-1643 ◽  
Author(s):  
S C Ho ◽  
M Schindler ◽  
J L Wang
Keyword(s):  
Isoelectric Focusing ◽  
Bradyrhizobium Japonicum ◽  
Affinity Column ◽  
Carbohydrate Binding ◽  
Binding Affinities ◽  
Isolation And Characterization ◽  
Relative Binding ◽  
Mannose Derivatives ◽  
Derivatives Of

Extracts of Bradyrhizobium japonicum were fractionated on Sepharose columns covalently derivatized with lactose. Elution of the material that was specifically bound to the affinity column with lactose yielded a protein of Mr approximately 38,000. Isoelectric focusing of this sample yielded two spots with pI values of 6.4 and 6.8. This protein specifically bound to galactose-containing glycoconjugates, but did not bind either to glucose or mannose. Derivatives of galactose at the C-2 position showed much weaker binding; there was an 18-fold difference in the relative binding affinities of galactose versus N-acetyl-D-galactosamine. These results indicate that we have purified a newly identified carbohydrate-binding protein from Bradyrhizobium japonicum, that can exquisitely distinguish galactose from its derivatives at the C-2 position.

scholarly journals Immunochemical characterization of human plasma fibronectin

1980 ◽  
Vol 191 (3) ◽  
pp. 719-727 ◽  
Author(s):  
M Vuento ◽  
E Salonen ◽  
K Salminen ◽  
M Pasanen ◽  
U K Stenman
Keyword(s):  
Human Plasma ◽  
Antigenic Structure ◽  
Antigenic Determinants ◽  
Proteolytic Degradation ◽  
Plasma Fibronectin ◽  
Native Protein ◽  
Covalent Interaction ◽  
Antigenic Activity ◽  
Haemagglutinating Activity

Human plasma fibronectin has been purified by a non-denaturing affinity chromatography procedure [Vuento & Vaheri, (1979) Biochem.J. 183, 331–337], and antisera have been raised by immunizing rabbits with the native protein. The antisera reacted strongly with native fibronectin, but only weakly with reduced and alkylated fibronectin or with heat-denaturated fibronectin. Denaturation also affected the haemagglutinating and gelatin-binding activities of fibronectin and increased its susceptibility to proteolytic degradation. The antisera reacted with fragments of fibronectin obtained by proteolysis with plasmin. Large fragments (mol.wt. 180000–200000), lacking the region harbouring the interchain disulphide bridges but containing the sites responsible for gelatin-binding and haemagglutinating activity, showed as intense a reaction with the antisera as intact fibronectin. Smaller peptides showed a weaker reaction. All fragments tested showed sensitivity to denaturation in their reaction with the antisera. The results were interpreted as showing that: (1) native fibronectin has an ordered conformation that is easily perturbed by denaturation; (2) most of the antigenic determinants of the protein are dependent on conformation; (3) the region of the fibronectin molecule containing the interchain disulphide bridges has only few antigenic determinants; and (4) covalent interaction of the two subunits does not contribute to the antigenic structure recognized by rabbit antisera. The observed correlation between the antigenic activity and a structural and functional intactness of fibronectin suggests that the antibodies to native fibronectin could be used as a conformational probe in studies on this protein.

Isolation and characterization of collagen-binding domains from human von Willebrand factor

2009 ◽  
Vol 24 (3) ◽  
pp. 150-154 ◽  
Author(s):  
N. E. Sharapova ◽  
A. P. Kotnova ◽  
Z. M. Galushkina ◽  
N. N. Poletaeva ◽  
N. V. Lavrova ◽  
...  
Keyword(s):  
Von Willebrand Factor ◽  
Collagen Binding ◽  
Binding Domains ◽  
Isolation And Characterization ◽  
Von Willebrand ◽  
Willebrand Factor

scholarly journals Isolation and Characterization of a Novel Collagen-binding Protein fromStreptococcus pyogenesStrain 6414

1995 ◽  
Vol 270 (1) ◽  
pp. 347-353 ◽  
Author(s):  
Livia Visai ◽  
Silvia Bozzini ◽  
Giuseppe Raucci ◽  
Antonio Toniolo ◽  
Pietro Speziale
Keyword(s):  
Binding Protein ◽  
Collagen Binding ◽  
Isolation And Characterization

Isolation and characterization of some proteinase inhibitors from Phaseolus vulgaris var. nanus

1980 ◽  
Vol 171 (1) ◽  
pp. 28-34 ◽  
Author(s):  
Holger Gerstenberg ◽  
Hans-Dieter Belitz ◽  
J�rgen K. P. Weder
Keyword(s):  
Phaseolus Vulgaris ◽  
Proteinase Inhibitors ◽  
Isolation And Characterization

Isolation and characterization of rat plasma glandular kallikrein

1985 ◽  
Vol 34 (1) ◽  
pp. 51-56 ◽  
Author(s):  
Jaime Masferrer ◽  
Renato Albertini ◽  
Héctor R. Croxatto ◽  
Pilar García ◽  
Inés Pinto
Keyword(s):  
Rat Plasma ◽  
Glandular Kallikrein ◽  
Isolation And Characterization

scholarly journals Purification and characterization of a tetrameric α-macroglobulin proteinase inhibitor from the gastropod mollusc Biomphalaria glabrata

1996 ◽  
Vol 316 (3) ◽  
pp. 893-900 ◽  
Author(s):  
Randall C. BENDER ◽  
Christopher J. BAYNE
Keyword(s):  
Quaternary Structure ◽  
Proteinase Inhibitors ◽  
Cysteine Proteinase ◽  
Serine Proteinase ◽  
Biomphalaria Glabrata ◽  
Sedimentation Equilibrium ◽  
Flight Mass Spectrometry ◽  
Isolation And Characterization ◽  
Α2 Macroglobulin

The α-macroglobulin proteinase inhibitors (αMs) are a family of proteins with the unique ability to inhibit a broad spectrum of proteinases. Whereas monomeric, dimeric and tetrameric αMs have been identified in vertebrates, all invertebrate αMs characterized so far have been dimeric. This paper reports the isolation and characterization of a tetrameric αM from the tropical planorbid snail Biomphalaria glabrata. The sequence of 18 amino acids at the N-terminus indicates homology with other αMs. The subunit mass of approx. 200 kDa was determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and SDS/PAGE. The quaternary structure was determined by sedimentation equilibrium centrifugation and native pore-limit electrophoresis. Evidence for a thioester is provided by the fact that methylamine treatment prevents the autolytic cleavage of the snail αM subunit and results in the release of 4 mol of thiols per mol of snail αM. The snail αM inhibited the serine proteinase trypsin, the cysteine proteinase bromelain and the metalloproteinase thermolysin. The spectrum of proteinases inhibited, together with the demonstration of steric protection of the proteinase active site and a ‘slow to fast’ conformational change after reacting with trypsin, all suggest that the inhibitory mechanism of the snail αM is similar to the ‘trap mechanism’ of human α2-macroglobulin.

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