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Purification of human complement protein C5.

Published on Jan 1, 2014in Methods of Molecular Biology
· DOI :10.1007/978-1-62703-724-2_7
Lars Sottrup-Jensen51
Estimated H-index: 51
(AU: Aarhus University),
Gregers R. Andersen36
Estimated H-index: 36
(AU: Aarhus University)
Abstract
  • References (13)
  • Citations (4)
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References13
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#1Cosmin A. Tegla (UMD: University of Maryland, College Park)H-Index: 12
#2Cornelia Cudrici (UMD: University of Maryland, College Park)H-Index: 21
Last. Horea Rus (UMD: University of Maryland, College Park)H-Index: 36
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Complement system activation plays an important role in both innate and acquired immunity. Activation of the complement and the subsequent formation of C5b-9 channels (the membrane attack complex) on the cell membranes lead to cell death. However, when the number of channels assembled on the surface of nucleated cells is limited, sublytic C5b-9 can induce cell cycle progression by activating signal transduction pathways and transcription factors and inhibiting apoptosis. This induction by C5b-9 ...
168 CitationsSource
#1N.S. Laursen (AU: Aarhus University)H-Index: 6
#2Kasper R. Andersen (AU: Aarhus University)H-Index: 15
Last. Gregers R. Andersen (AU: Aarhus University)H-Index: 36
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Complement acts as a danger-sensing system in the innate immune system, and its activation initiates a strong inflammatory response and cleavage of the proteins C3 and C5 by proteolytic enzymes, the convertases. These contain a non-catalytic substrate contacting subunit (C3b or C4b) in complex with a protease subunit (Bb or C2a). We determined the crystal structures of the C3b homologue cobra venom factor (CVF) in complex with C5, and in complex with C5 and the inhibitor SSL7 at 4.3 A resolution...
55 CitationsSource
#1Daniel Ricklin (UPenn: University of Pennsylvania)H-Index: 39
#2George Hajishengallis (University of Louisville)H-Index: 57
Last. John D. Lambris (UPenn: University of Pennsylvania)H-Index: 95
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Nearly a century after the significance of the human complement system was recognized we have come to realize that its versatile functions extend far beyond the elimination of microbes. Indeed, complement acts as a rapid and efficient immune surveillance system that has distinct effects on healthy and altered host cells and foreign intruders. By eliminating cellular debris and infectious microbes, orchestrating immune responses, and sending `danger' signals, complement contributes substantially ...
1,797 CitationsSource
Staphylococcus aureus secretes the SSL7 protein as part of its immune evasion strategy. The protein binds both complement C5 and IgA, yet it is unclear whether SSL7 cross-links these two proteins and, if so, what purpose this serves the pathogen. We have isolated a stable IgA-SSL7-C5 complex, and our crystal structure of the C5-SSL7 complex confirms that binding to C5 occurs exclusively through the C-terminal β-grasp domain of SSL7 leaving the OB domain free to interact with IgA. SSL7 interacts ...
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#1Andreas KlosH-Index: 33
#2Andrea J. Tenner (UCI: University of California, Irvine)H-Index: 55
Last. Jörg Köhl (University of Lübeck)H-Index: 55
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The anaphylatoxins (AT) C3a, C5a and C5a-desArg are generally considered pro-inflammatory polypeptides generated after proteolytic cleavage of C3 and C5 in response to complement activation. Their well appreciated effector functions include chemotaxis and activation of granulocytes, mast cells and macrophages. Recent evidence suggests that ATs are also generated locally within tissues by pathogen-, cell-, or contact system-derived proteases. This local generation of ATs is important for their pl...
371 CitationsSource
#1Folmer Fredslund (AU: Aarhus University)H-Index: 10
#2N.S. Laursen (AU: Aarhus University)H-Index: 6
Last. Gregers R. Andersen (AU: Aarhus University)H-Index: 36
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To provide insight into the structural and functional properties of human complement component 5 (C5), we determined its crystal structure at a resolution of 3.1 A. The core of C5 adopted a structure resembling that of C3, with the domain arrangement at the position corresponding to the C3 thioester being very well conserved. However, in contrast to C3, the convertase cleavage site in C5 was ordered and the C345C domain flexibly attached to the core of C5. Binding of the tick C5 inhibitor OmCI t...
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#1M. K. Pangburn (University of Texas Health Science Center at San Antonio)H-Index: 2
#2N. Rawal (University of Texas Health Science Center at San Antonio)H-Index: 2
The multisubunit enzymes of the complement system that cleave C5 have many unusual properties, the most striking of which is that they acquire their specificity for C5 following cleavage of another substrate C3. C5 convertases are assemblies of two proteins C4b and C2a (classical or lectin pathways) or C3b and Bb (alternative pathway) and additional C3b molecules. The catalytic complexes (C4b, C2a or C3b, Bb) are intrinsically unstable ( t 1,2 = 1–3 min) and the enzymes are controlled by numerou...
88 CitationsSource
The molecular mechanisms of the alternative pathway are now reasonably well understood. Intramolecular functional sites are currently being delineated that govern the crucial interactions between C3 and the other proteins of the pathway. The knowledge of the primary structure of C3 is affording the synthesis of small peptides that may aid the identification of functional regions. Should the three-dimensional structure of C3 or C3b become available, information on functional regions may readily b...
106 CitationsSource
#1Lars Sottrup-Jensen (AU: Aarhus University)H-Index: 51
113 CitationsSource
#1Carl H. HammerH-Index: 35
#2G H WirtzH-Index: 1
Last. B F TackH-Index: 1
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Abstract In the present work a scheme is presented for the isolation of multiple components of human complement in a functionally and biochemically pure state and with full hemolytic activity. These preparative procedures allow high recovery of milligram and gram quantities of particular complement components from a large pool (2-11 liters) of fresh EDTA plasma in no more than four chromatographic steps. Many components (C3bINA, C5, C3, C1EI, C4, and C9) are recovered functionally pure or highly...
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#1Shutong Xu (Harvard University)H-Index: 2
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Recognition by the leukocyte integrins α X β 2 and α M β 2 of complement iC3b-opsonized targets is essential for effector functions including phagocytosis. The integrin-binding sites on iC3b remain incompletely characterized. Here, we describe negative-stain electron microscopy and biochemical studies of α X β 2 and α M β 2 in complex with iC3b. Despite high homology, the two integrins bind iC3b at multiple distinct sites. α X β 2 uses the α X αI domain to bind iC3b on its C3c moiety at one of t...
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Eculizumab is a humanized mAb approved for treatment of patients with paroxysmal nocturnal hemoglobinuria and atypical hemolytic uremic syndrome. Eculizumab binds complement component C5 and prevents its cleavage by C5 convertases, inhibiting release of both the proinflammatory metabolite C5a and formation of the membrane attack complex via C5b. In this study, we present the crystal structure of the complex between C5 and a Fab fragment with the same sequence as eculizumab at a resolution of 4.2...
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