Purification of human plasma fibronectin using immobilized gelatin and Arg affinity chromatography

2008 ◽  
Vol 3 (3) ◽  
pp. 525-533 ◽  
Author(s):  
Pietro Speziale ◽  
Livia Visai ◽  
Simonetta Rindi ◽  
Antonella Di Poto
Keyword(s):  
Affinity Chromatography ◽  
Human Plasma ◽  
Plasma Fibronectin

scholarly journals Primary structure of a glycosylated DNA-binding domain in human plasma fibronectin.

1985 ◽  
Vol 260 (4) ◽  
pp. 2301-2306
Author(s):  
H Pande ◽  
J Calaycay ◽  
D Hawke ◽  
C M Ben-Avram ◽  
J E Shively
Keyword(s):  
Dna Binding ◽  
Human Plasma ◽  
Primary Structure ◽  
Binding Domain ◽  
Dna Binding Domain ◽  
Plasma Fibronectin

scholarly journals Primary structure of human plasma fibronectin. The 29,000-dalton NH2-terminal domain.

1983 ◽  
Vol 258 (20) ◽  
pp. 12670-12674 ◽  
Author(s):  
A Garcia-Pardo ◽  
E Pearlstein ◽  
B Frangione
Keyword(s):  
Human Plasma ◽  
Primary Structure ◽  
Plasma Fibronectin ◽  
Terminal Domain

scholarly journals Thermal stability of human plasma fibronectin and its constituent domains.

1984 ◽  
Vol 259 (19) ◽  
pp. 11901-11907 ◽  
Author(s):  
K C Ingham ◽  
S A Brew ◽  
T J Broekelmann ◽  
J A McDonald
Keyword(s):  
Thermal Stability ◽  
Human Plasma ◽  
Plasma Fibronectin ◽  
Thermal Stability Of

scholarly journals Human plasma fibronectin as a substrate for human urokinase

1989 ◽  
Vol 262 (2) ◽  
pp. 529-534 ◽  
Author(s):  
L I Gold ◽  
R Schwimmer ◽  
J P Quigley
Keyword(s):  
Extracellular Matrix ◽  
Human Plasma ◽  
Plasminogen Activator ◽  
Early Event ◽  
Plasma Fibronectin ◽  
Proteolytic Digestion ◽  
Enzyme Substrate ◽  
Tumour Cell Invasion ◽  
Substrate Ratio ◽  
Dose Dependent

An early event in malignant transformation is the increased expression of proteases, such as plasminogen activator, which can degrade surrounding extracellular matrices, thereby conferring an advantage for tumour cell invasion and metastasis. The present studies provide evidence that plasma fibronectin (Fn), which is a component of the extracellular matrix, is a direct substrate for the plasminogen activator urokinase (UK). Human plasma Fn was incubated with human UK under plasminogen-free conditions. Fn cleavage was both time- and dose-dependent and was evident within 30 min. The proteolytic digestion was limited and complete within 12 h at an enzyme/substrate ratio of 1:20. Analysis of the final proteolytic digestion products demonstrated the disappearance of the native dimeric 440 kDa structure of Fn with the concomitant appearance of three proteolytic fragments of 210, 200 and 25 kDa. Since two large fragments of similar size to the 220 kDa monomeric chains of Fn were obtained following proteolysis, it is proposed that UK cleaves Fn at two sites, one towards the N-terminal and one close to the C-terminal, but N-terminal to its interchain disulphide bonds. These studies suggest that the local proteolytic digestion and release of Fn from the extracellular matrix by tumour cells possessing high levels of UK may involve the direct proteolytic breakdown of Fn by UK.

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.

Direct analysis – no sample preparation – of bioavailable cortisol in human plasma by weak affinity chromatography (WAC)

2017 ◽  
Vol 1061-1062 ◽  
pp. 438-444 ◽  
Author(s):  
Sten Ohlson ◽  
Jagjit Kaur ◽  
Manfred Raida ◽  
Ulf Niss ◽  
Tim Bengala ◽  
...  
Keyword(s):  
Sample Preparation ◽  
Affinity Chromatography ◽  
Human Plasma ◽  
Direct Analysis ◽  
No Sample Preparation
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