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Isolation and chara...
Isolation and characterization of hemolymph clotting factors in Drosophila melanogaster by a pullout method
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Scherfer, C (författare)
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Karlsson, C (författare)
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- Loseva, O. (författare)
- Stockholms universitet,Institutionen för genetik, mikrobiologi och toxikologi
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Bidla, G (författare)
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Goto, A (författare)
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Havemann, J (författare)
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- Dushay, Mitchell S (författare)
- Södertörns högskola,Institutionen för kemi, biologi, geografi och miljövetenskap
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- Theopold, U. (författare)
- Stockholms universitet,Institutionen för molekylärbiologi och funktionsgenomik
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- Wilhelmsson, C. (författare)
- Stockholms universitet,Institutionen för molekylärbiologi och funktionsgenomik
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(creator_code:org_t)
- Elsevier BV, 2004
- 2004
- Engelska.
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Ingår i: Current Biology. - : Elsevier BV. - 0960-9822 .- 1879-0445. ; 14:7, s. 625-629
- Relaterad länk:
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http://www.cell.com/...
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https://urn.kb.se/re...
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https://doi.org/10.1...
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https://urn.kb.se/re...
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Abstract
Ämnesord
Stäng
- Clotting is critical in limiting loss of hemolymph and initiating wound healing in insects as well as in vertebrates [1]. Clotting is also an important immune defense, quickly forming a secondary barrier to infection, thereby immobilizing, and possibly killing bacteria directly [2, 3]. Here, we describe methods to assess clotting and to extract the clot from Drosophila larval hemolymph by using aggregation of paramagnetic beads. The validity of the assay was demonstrated by characterization of mutants. We show that clotting occurs in the absence of phenoloxidase and that the Drosophila clot binds bacteria. We also describe a pullout assay to purify the clot as a whole, free from entrapped hemocytes and cellular debris. Proteins subsequently identified by mass spectrometry include both predicted and novel clot proteins. Immune induction has been shown for three of the latter, namely Tiggrin and two unknown proteins (GC15825 and CG15293) [4,5] that we now propose function in hemolymph clotting. The most abundant clot protein is Hemolectin [6], and we confirm that hemolectin mutant larvae show clotting defects.
Ämnesord
- NATURVETENSKAP -- Biologi -- Cellbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Cell Biology (hsv//eng)
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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