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  Fluorescence microscopy visualization of halomucin, a secreted 927 kDa protein surrounding Haloquadratum walsbyi cells

Zenke, R., von Gronau, S., Bolhuis, H., Gruska, M., Pfeiffer, F., & Oesterhelt, D. (2015). Fluorescence microscopy visualization of halomucin, a secreted 927 kDa protein surrounding Haloquadratum walsbyi cells. FRONTIERS IN MICROBIOLOGY, 6: 249. doi:10.3389/fmicb.2015.00249.

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 Creators:
Zenke, Ralf1, Author           
von Gronau, Susanne2, Author           
Bolhuis, Henk3, Author
Gruska, Manuela3, Author
Pfeiffer, Friedhelm2, Author           
Oesterhelt, Dieter2, Author           
Affiliations:
1Scientific Service Groups, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565170              
2Oesterhelt, Dieter / Membrane Biochemistry, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565164              
3external, ou_persistent22              

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Free keywords: SQUARE HALOARCHAEON; BRINE POOL; SP NOV.; BACTERIA; CULTIVATION; ARCHAEON; SINAIhalomucin; halophilic archaea; polyhydroxybutyrate; cell shape; Haloquadratum; immunofluorescence; protein secretion;
 Abstract: At the time of its first publication, halomucin from Haloquadratum walsbyi strain HBSQ001 was the largest archaeal protein known (9159 aa). It has a predicted signal sequence, making it likely to be an extracellular or secreted protein. Best BLAST matches were found to be mammalian mucins that protect tissues to dehydration and chemical stress. It was hypothesized that halomucin participates in protection against desiccation by retaining water in a hull around the halophilic organisms that live at the limits of water activity. We visualized Halo quadratum cells by staining their intracellular polyhydroxybutyrate granules using Nile Blue. Halomucin was stained by immunofluorescence with antibodies generated against synthetic peptides derived from the halomucin amino acid sequence. Polyhydroxybutyrate stained cells were reconstructed in 3D which highlights not only the highly regular square shape but also the extreme flatness of Haloquadratum. Double-staining proves halomucin to be extracellular but to be only loosely associated to cells in agreement with its hypothesized function.

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Language(s): eng - English
 Dates: 2015
 Publication Status: Published online
 Pages: 6
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: ISI: 000352536400001
DOI: 10.3389/fmicb.2015.00249
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Title: FRONTIERS IN MICROBIOLOGY
Source Genre: Journal
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Publ. Info: PO BOX 110, LAUSANNE, 1015, SWITZERLAND : FRONTIERS RESEARCH FOUNDATION
Pages: - Volume / Issue: 6 Sequence Number: 249 Start / End Page: - Identifier: ISSN: 1664-302X