This article was downloaded by: [Rutgers University] On: 04 April 2015, At: 10:06 Publisher: Taylor & Francis Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered office: Mortimer House, 37-41 Mortimer Street, London W1T 3JH, UK
Journal of Biomolecular Structure and Dynamics Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/tbsd20
Dynamic Light Scattering Studies on Hydrodynamic Properties of FibrinogenFibronectin Complex a
b
Kazuyuki Nagamatsu , Masaki Komori , Shigeomi b
Kuroda & Koji Tanaka
a
a
Department of Materials Science and Technology , Yamaguchi College Science University of Tokyo , Onoda , Yamaguchi , 756 , JAPAN b
Internal Medicine Ohkura National Hospital , Setagaya-ku , Tokyo , JAPAN Published online: 21 May 2012.
To cite this article: Kazuyuki Nagamatsu , Masaki Komori , Shigeomi Kuroda & Koji Tanaka (1992) Dynamic Light Scattering Studies on Hydrodynamic Properties of Fibrinogen-Fibronectin Complex, Journal of Biomolecular Structure and Dynamics, 9:4, 807-820, DOI: 10.1080/07391102.1992.10507957 To link to this article: http://dx.doi.org/10.1080/07391102.1992.10507957
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages,
and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content.
Downloaded by [Rutgers University] at 10:06 04 April 2015
This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/terms-and-conditions
Journal of Biomolecular Structure & Dynamics, JSSN 0739-1102 Volume 9, Issue Number 4 (1992), ®Adenine Press (1992).
Dynamic Light Scattering Studies on Hydrodynamic Properties of Fibrinogen-Fibronectin Complex
Downloaded by [Rutgers University] at 10:06 04 April 2015
Kazuyuki Nagamatsu 1, Masaki Komori2 , Shigeomi Kuroda 2 and Koji Tanaka 1 'Department of Materials Science and Technology Yamaguchi College Science University of Tokyo Onoda, Yamaguchi 756, JAPAN 2
Intemal Medicine Ohkura National Hospital Setagaya-ku, Tokyo, JAPAN Abstract A high molecular weight' cryogel' was obtained as insoluble complexes by cold incubation at near-freezing temperatures from heparinized plasma of patients with rheumatoid arthritis. After the cryogel was solubilized at 37 °C,l: 1 complex of fibrinogen and fibronectin was purified at room temperature by affinity chromatography on a gelatin-Sepharose 4B. Hydrodynamic properties of the complex were investigated as a function of temperature and NaCl concentration using a dynamic light scattering. The diffusion coefficients of the complex at 20 ac decreased with increasing ofNaCl concentration as free fibronectin. The complex appears to be a more compact form at low ionic concentration, which is associated with conformational changes of fibronectin. The diffusion coefficient of the complex at 20 ac in 0.05 M TrisHCl(pH7.4) containing 0.5 M NaCl was estimated as 8.5xlo-s cm2s- 1• The complex did not dissociate over the temperature range from 20 to 37 a c. The diffusion coefficients of the complex decreased significantly at 12 ac and 40 ac. The thermal denaturation of fibrinogen molecule in the complex was observed at40 a c. The CONTIN analysis of the light scattering data showed that the complex associated to form higher aggregates at 15 a c. but not at nearfreezing temperature. The equilibrium between the complex and higher aggregates appeared reversible.
Introduction In 1954, Thomas eta/. described the formation of plasma precipitate induced in the cold by heparin( 1). The cryoprecipitates were found more abundantly in plasma of patients with various inflammatory, infectious, or neoplastic diseases(2,3). Smith and von Korff speculated that the components of the cryoprecipitates are fibrinogen and cold insoluble globulin(Cig) which later was termed 'fibronectin'. Subsequent studies confirmed their speculations from analysis of the cryoprecipitates obtained from patients with dermatological diseases and ulcers(4,5). However, it is not clear
807
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Nagamatsu et a/.
Downloaded by [Rutgers University] at 10:06 04 April 2015
as to why the cryoprecipitates are induced from such patients to larger contents. Smith and von Korff investigated effects of temperature, ionic strength and pH on the occurrence of cryoprecipitates(2,3). Stathukis and Mosesson clarified the mechanism of cryoprecipitate formation and suggested that fibrinogen associated with a fibronectincontaining nucleus(6). Their studies is experimentally demonstrated by turbidity measurements of cryoprecipitates formed at near-freezing temperatures. In recent years, plasmapheresis has been practiced on patients with rheumatoid arthritis(RA) and its clinical effectiveness has been reported(7,8). Acryofiltration(CF) method for plasmapheresis applies that a high molecular weight cryogel is formed by the cold incubation of heparinized plasma(9). We obtained removal substances, so-called cryogel, by the CF method from patients with RA, and reported analysis of cryogel by immunoelectrophoresis and high performance liquid chromatography and clinical significance(lO). To study physicochemical properties of the complex between fibrinogen and fibronectin, a method for purification of the component from cryogel with high yield is important. We used an affinity chromatography on gelatin-Sepharose 4B, which has been used for isolation of fibronectin from plasma, and purified 1:1 complex of fibrinogen and fibronectin. Much information concerning the structure and functions of fibrinogen and fibronectin has been reported( 11-16). Electron microscope images demonstrated the basic structure of the molecule: a long, thin, cylindrical strand. Fibrinogen and fibronectin appear to be basically the similar structure. On the other hand, dynamic light scattering method has been utilized to yield solution properties ofbiomolecules{l7). Under the slightly different conditions, comparable values of 2.0X 10- 7 cm2s- 1 for fibrinogen and 2.1X 10- 7 cm2s - 1 for fibronectin were reported(l8-21). Few physical studies of interactions between fibrinogen and fibronectin have been reported. In this paper, we used dynamic light scattering to monitor hydrodynamic properties of the complex and investigated the equilibrium between the complex and higher aggregates as a function of temperature and ionic concentration.
Materials and Methods All reagents used were of analytical grade and obtained from commercial suppliers unless otherwise indicated. All solutions used for analysis were filtered through 0.22 11m filters(Sterivex-GS, Millipore). The CF system was performed as described previously(22,23). Plasma separated by a first filter was cooled at 4 °C, and then colloidal precipitate was eliminated by using a second filter. An AP-05H (As a hi Medical, Japan) and a PS-4000 (Terumo, Japan) were used as the first filter, and an AP-06 (Asahi Medical) and CF-01 (Terumo) as the second filter. When the pressure in the second filter rose to approximately 250 mmHg, CF was terminated. Cryofiltration was performed 486 times on 29 patients with RA. The plasma in the circuit was returned to the patients. The cryogel in the second filter was flushed out into test tubes. The flushed solution was kept in a deep freezer( -70 oq until analysis. Analysis of cryogel by high performance liquid chromatography(HPLC) was performed
809
Fibrinogen·Fibronectin Complex
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Journal of Biomolecular Structure and Dynamics Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/tbsd20
Dynamic Light Scattering Studies on Hydrodynamic Properties of FibrinogenFibronectin Complex a
b
Kazuyuki Nagamatsu , Masaki Komori , Shigeomi b
Kuroda & Koji Tanaka
a
a
Department of Materials Science and Technology , Yamaguchi College Science University of Tokyo , Onoda , Yamaguchi , 756 , JAPAN b
Internal Medicine Ohkura National Hospital , Setagaya-ku , Tokyo , JAPAN Published online: 21 May 2012.
To cite this article: Kazuyuki Nagamatsu , Masaki Komori , Shigeomi Kuroda & Koji Tanaka (1992) Dynamic Light Scattering Studies on Hydrodynamic Properties of Fibrinogen-Fibronectin Complex, Journal of Biomolecular Structure and Dynamics, 9:4, 807-820, DOI: 10.1080/07391102.1992.10507957 To link to this article: http://dx.doi.org/10.1080/07391102.1992.10507957
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages,
and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content.
Downloaded by [Rutgers University] at 10:06 04 April 2015
This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/terms-and-conditions
Journal of Biomolecular Structure & Dynamics, JSSN 0739-1102 Volume 9, Issue Number 4 (1992), ®Adenine Press (1992).
Dynamic Light Scattering Studies on Hydrodynamic Properties of Fibrinogen-Fibronectin Complex
Downloaded by [Rutgers University] at 10:06 04 April 2015
Kazuyuki Nagamatsu 1, Masaki Komori2 , Shigeomi Kuroda 2 and Koji Tanaka 1 'Department of Materials Science and Technology Yamaguchi College Science University of Tokyo Onoda, Yamaguchi 756, JAPAN 2
Intemal Medicine Ohkura National Hospital Setagaya-ku, Tokyo, JAPAN Abstract A high molecular weight' cryogel' was obtained as insoluble complexes by cold incubation at near-freezing temperatures from heparinized plasma of patients with rheumatoid arthritis. After the cryogel was solubilized at 37 °C,l: 1 complex of fibrinogen and fibronectin was purified at room temperature by affinity chromatography on a gelatin-Sepharose 4B. Hydrodynamic properties of the complex were investigated as a function of temperature and NaCl concentration using a dynamic light scattering. The diffusion coefficients of the complex at 20 ac decreased with increasing ofNaCl concentration as free fibronectin. The complex appears to be a more compact form at low ionic concentration, which is associated with conformational changes of fibronectin. The diffusion coefficient of the complex at 20 ac in 0.05 M TrisHCl(pH7.4) containing 0.5 M NaCl was estimated as 8.5xlo-s cm2s- 1• The complex did not dissociate over the temperature range from 20 to 37 a c. The diffusion coefficients of the complex decreased significantly at 12 ac and 40 ac. The thermal denaturation of fibrinogen molecule in the complex was observed at40 a c. The CONTIN analysis of the light scattering data showed that the complex associated to form higher aggregates at 15 a c. but not at nearfreezing temperature. The equilibrium between the complex and higher aggregates appeared reversible.
Introduction In 1954, Thomas eta/. described the formation of plasma precipitate induced in the cold by heparin( 1). The cryoprecipitates were found more abundantly in plasma of patients with various inflammatory, infectious, or neoplastic diseases(2,3). Smith and von Korff speculated that the components of the cryoprecipitates are fibrinogen and cold insoluble globulin(Cig) which later was termed 'fibronectin'. Subsequent studies confirmed their speculations from analysis of the cryoprecipitates obtained from patients with dermatological diseases and ulcers(4,5). However, it is not clear
807
BOB
Nagamatsu et a/.
Downloaded by [Rutgers University] at 10:06 04 April 2015
as to why the cryoprecipitates are induced from such patients to larger contents. Smith and von Korff investigated effects of temperature, ionic strength and pH on the occurrence of cryoprecipitates(2,3). Stathukis and Mosesson clarified the mechanism of cryoprecipitate formation and suggested that fibrinogen associated with a fibronectincontaining nucleus(6). Their studies is experimentally demonstrated by turbidity measurements of cryoprecipitates formed at near-freezing temperatures. In recent years, plasmapheresis has been practiced on patients with rheumatoid arthritis(RA) and its clinical effectiveness has been reported(7,8). Acryofiltration(CF) method for plasmapheresis applies that a high molecular weight cryogel is formed by the cold incubation of heparinized plasma(9). We obtained removal substances, so-called cryogel, by the CF method from patients with RA, and reported analysis of cryogel by immunoelectrophoresis and high performance liquid chromatography and clinical significance(lO). To study physicochemical properties of the complex between fibrinogen and fibronectin, a method for purification of the component from cryogel with high yield is important. We used an affinity chromatography on gelatin-Sepharose 4B, which has been used for isolation of fibronectin from plasma, and purified 1:1 complex of fibrinogen and fibronectin. Much information concerning the structure and functions of fibrinogen and fibronectin has been reported( 11-16). Electron microscope images demonstrated the basic structure of the molecule: a long, thin, cylindrical strand. Fibrinogen and fibronectin appear to be basically the similar structure. On the other hand, dynamic light scattering method has been utilized to yield solution properties ofbiomolecules{l7). Under the slightly different conditions, comparable values of 2.0X 10- 7 cm2s- 1 for fibrinogen and 2.1X 10- 7 cm2s - 1 for fibronectin were reported(l8-21). Few physical studies of interactions between fibrinogen and fibronectin have been reported. In this paper, we used dynamic light scattering to monitor hydrodynamic properties of the complex and investigated the equilibrium between the complex and higher aggregates as a function of temperature and ionic concentration.
Materials and Methods All reagents used were of analytical grade and obtained from commercial suppliers unless otherwise indicated. All solutions used for analysis were filtered through 0.22 11m filters(Sterivex-GS, Millipore). The CF system was performed as described previously(22,23). Plasma separated by a first filter was cooled at 4 °C, and then colloidal precipitate was eliminated by using a second filter. An AP-05H (As a hi Medical, Japan) and a PS-4000 (Terumo, Japan) were used as the first filter, and an AP-06 (Asahi Medical) and CF-01 (Terumo) as the second filter. When the pressure in the second filter rose to approximately 250 mmHg, CF was terminated. Cryofiltration was performed 486 times on 29 patients with RA. The plasma in the circuit was returned to the patients. The cryogel in the second filter was flushed out into test tubes. The flushed solution was kept in a deep freezer( -70 oq until analysis. Analysis of cryogel by high performance liquid chromatography(HPLC) was performed
809
Fibrinogen·Fibronectin Complex
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