Hydrodynamic and mass spectrometry analysis of nearly-intact human fibrinogen, chicken fibrinogen, and of a substantially monodisperse human fibrinogen fragment X

Barbara Cardinali, Aldo Profumo, Anna Aprile, Olwyn Byron, Gordon Morris, Stephen E. Harding, Walter F. Stafford, Mattia Rocco

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

The shape and solution properties of fibrinogen are affected by the location of the C-terminal portion of the Aα chains, which is presently still controversial. We have measured the hydrodynamic properties of a human fibrinogen fraction with these appendages mostly intact, of chicken fibrinogen, where they lack 11 characteristic 13-amino acids repeats, and of human fragment X, a plasmin early degradation product in which they have been removed. The human fibrinogen/fragment X samples were extensively characterized by SDS-PAGE/Western blotting and mass spectrometry, allowing their composition to be precisely determined. The solution properties of all samples were then investigated by analytical ultracentrifugation and size-exclusion HPLC coupled with multi-angle light scattering and differential pressure viscometry detectors. The measured parameters suggest that the extra repeats have little influence on the overall fibrinogen conformation, while a significant change is brought about by the removal of the C-terminal portion of the Aα chains beyond residue Aα200.

Original languageEnglish
Pages (from-to)157-168
Number of pages12
JournalArchives of Biochemistry and Biophysics
Volume493
Issue number2
Early online date22 Oct 2009
DOIs
Publication statusPublished - 15 Jan 2010
Externally publishedYes

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Hydrodynamics
Fibrinogen
Mass spectrometry
Chickens
Mass Spectrometry
Fibrinolysin
Ultracentrifugation
Viscosity measurement
Light scattering
Conformations
Polyacrylamide Gel Electrophoresis
Western Blotting
High Pressure Liquid Chromatography
Detectors
Light
Amino Acids
Pressure
Degradation
fibrinogen fragment X
Chemical analysis

Cite this

Cardinali, Barbara ; Profumo, Aldo ; Aprile, Anna ; Byron, Olwyn ; Morris, Gordon ; Harding, Stephen E. ; Stafford, Walter F. ; Rocco, Mattia. / Hydrodynamic and mass spectrometry analysis of nearly-intact human fibrinogen, chicken fibrinogen, and of a substantially monodisperse human fibrinogen fragment X. In: Archives of Biochemistry and Biophysics. 2010 ; Vol. 493, No. 2. pp. 157-168.
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Hydrodynamic and mass spectrometry analysis of nearly-intact human fibrinogen, chicken fibrinogen, and of a substantially monodisperse human fibrinogen fragment X. / Cardinali, Barbara; Profumo, Aldo; Aprile, Anna; Byron, Olwyn; Morris, Gordon; Harding, Stephen E.; Stafford, Walter F.; Rocco, Mattia.

In: Archives of Biochemistry and Biophysics, Vol. 493, No. 2, 15.01.2010, p. 157-168.

Research output: Contribution to journalArticle

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AU - Cardinali, Barbara

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AU - Aprile, Anna

AU - Byron, Olwyn

AU - Morris, Gordon

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AU - Stafford, Walter F.

AU - Rocco, Mattia

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AB - The shape and solution properties of fibrinogen are affected by the location of the C-terminal portion of the Aα chains, which is presently still controversial. We have measured the hydrodynamic properties of a human fibrinogen fraction with these appendages mostly intact, of chicken fibrinogen, where they lack 11 characteristic 13-amino acids repeats, and of human fragment X, a plasmin early degradation product in which they have been removed. The human fibrinogen/fragment X samples were extensively characterized by SDS-PAGE/Western blotting and mass spectrometry, allowing their composition to be precisely determined. The solution properties of all samples were then investigated by analytical ultracentrifugation and size-exclusion HPLC coupled with multi-angle light scattering and differential pressure viscometry detectors. The measured parameters suggest that the extra repeats have little influence on the overall fibrinogen conformation, while a significant change is brought about by the removal of the C-terminal portion of the Aα chains beyond residue Aα200.

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