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The Origin and Evolution of the Plant Cell Surface: Algal Integrin-Associated Proteins and a New Family of Integrin-Like Cytoskeleton-ECM Linker Proteins.

Becker B, Doan JM, Wustman B, Carpenter EJ, Chen L, Zhang Y, Wong GK, Melkonian M - Genome Biol Evol (2015)

Bottom Line: The extracellular matrix of scaly green flagellates consists of small organic scales consisting of polysaccharides and scale-associated proteins (SAPs).Gralins are absent from embryophytes.A model for the evolution of the cell surface proteins in Plantae is discussed.

View Article: PubMed Central - PubMed

Affiliation: Biozentrum Köln, Botanical Institute, Universität zu Köln, Germany b.becker@uni-koeln.de.

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SDS-PAGE and Western blot of isolated flagellar scales from Scherffelia dubia probed with anti-SAP98. (A) Coomassie Brilliant Blue stain of an isolated flagellar scale fraction (sf) run with a 12% SDS-PAGE. Circle, SAP98. The other SAPs for which peptide sequences were obtained are indicated by arrow heads. (B) Western blot of the same flagellar scale fraction probed with a polyclonal anti-SAP98 (8% SDS-PAGE). Lane 1 flagellar scale fraction after deglycosylation with N-Glycosidase F and lane 2 isolated flagellar scale fraction. The position of the prestained marker proteins is indicated in between both PAGE.
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evv089-F2: SDS-PAGE and Western blot of isolated flagellar scales from Scherffelia dubia probed with anti-SAP98. (A) Coomassie Brilliant Blue stain of an isolated flagellar scale fraction (sf) run with a 12% SDS-PAGE. Circle, SAP98. The other SAPs for which peptide sequences were obtained are indicated by arrow heads. (B) Western blot of the same flagellar scale fraction probed with a polyclonal anti-SAP98 (8% SDS-PAGE). Lane 1 flagellar scale fraction after deglycosylation with N-Glycosidase F and lane 2 isolated flagellar scale fraction. The position of the prestained marker proteins is indicated in between both PAGE.

Mentions: The subcellular localization of SAP126, a member of the large high-molecular weight complex (>2MDa) is known (Becker et al. 1996); however, the localization of the second SAP complex of 190 kDa, which includes SAP98, has not yet been determined. To address this question, the C-terminal part of SAP98 (SAP98-C, 484 amino acids) was expressed in bacteria (not shown). SAP98-C was purified by SDS-PAGE from inclusion bodies and subsequently used for preparation of a polyclonal antibody. On Western blots (fig. 2), the antibody recognized a protein of 98 kDa (80 kDa after deglycosylation with N-Glykosidase F). The apparent molecular mass of the deglycosylated protein (SDS-PAGE) is in very good agreement with the predicted molecular size (79.1 kDa) for SAP98. Immunofluorescence revealed that SAP98 is localized to the flagella (fig. 3). Often the fluorescence was located in two clearly distinguishable rows (fig. 3), suggesting that SAP98 may localize to the basal part of the flagellar hair scales, which are arranged in two opposite rows perpendicular to the plane of flagellar beat (fig. 4A). Attempts to localize the protein at the ultrastructural level were unsuccessful.Fig. 2.—


The Origin and Evolution of the Plant Cell Surface: Algal Integrin-Associated Proteins and a New Family of Integrin-Like Cytoskeleton-ECM Linker Proteins.

Becker B, Doan JM, Wustman B, Carpenter EJ, Chen L, Zhang Y, Wong GK, Melkonian M - Genome Biol Evol (2015)

SDS-PAGE and Western blot of isolated flagellar scales from Scherffelia dubia probed with anti-SAP98. (A) Coomassie Brilliant Blue stain of an isolated flagellar scale fraction (sf) run with a 12% SDS-PAGE. Circle, SAP98. The other SAPs for which peptide sequences were obtained are indicated by arrow heads. (B) Western blot of the same flagellar scale fraction probed with a polyclonal anti-SAP98 (8% SDS-PAGE). Lane 1 flagellar scale fraction after deglycosylation with N-Glycosidase F and lane 2 isolated flagellar scale fraction. The position of the prestained marker proteins is indicated in between both PAGE.
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Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4494055&req=5

evv089-F2: SDS-PAGE and Western blot of isolated flagellar scales from Scherffelia dubia probed with anti-SAP98. (A) Coomassie Brilliant Blue stain of an isolated flagellar scale fraction (sf) run with a 12% SDS-PAGE. Circle, SAP98. The other SAPs for which peptide sequences were obtained are indicated by arrow heads. (B) Western blot of the same flagellar scale fraction probed with a polyclonal anti-SAP98 (8% SDS-PAGE). Lane 1 flagellar scale fraction after deglycosylation with N-Glycosidase F and lane 2 isolated flagellar scale fraction. The position of the prestained marker proteins is indicated in between both PAGE.
Mentions: The subcellular localization of SAP126, a member of the large high-molecular weight complex (>2MDa) is known (Becker et al. 1996); however, the localization of the second SAP complex of 190 kDa, which includes SAP98, has not yet been determined. To address this question, the C-terminal part of SAP98 (SAP98-C, 484 amino acids) was expressed in bacteria (not shown). SAP98-C was purified by SDS-PAGE from inclusion bodies and subsequently used for preparation of a polyclonal antibody. On Western blots (fig. 2), the antibody recognized a protein of 98 kDa (80 kDa after deglycosylation with N-Glykosidase F). The apparent molecular mass of the deglycosylated protein (SDS-PAGE) is in very good agreement with the predicted molecular size (79.1 kDa) for SAP98. Immunofluorescence revealed that SAP98 is localized to the flagella (fig. 3). Often the fluorescence was located in two clearly distinguishable rows (fig. 3), suggesting that SAP98 may localize to the basal part of the flagellar hair scales, which are arranged in two opposite rows perpendicular to the plane of flagellar beat (fig. 4A). Attempts to localize the protein at the ultrastructural level were unsuccessful.Fig. 2.—

Bottom Line: The extracellular matrix of scaly green flagellates consists of small organic scales consisting of polysaccharides and scale-associated proteins (SAPs).Gralins are absent from embryophytes.A model for the evolution of the cell surface proteins in Plantae is discussed.

View Article: PubMed Central - PubMed

Affiliation: Biozentrum Köln, Botanical Institute, Universität zu Köln, Germany b.becker@uni-koeln.de.

Show MeSH