Action of Saponin on Biological Cell Membranes

Bangham, A. D.; Horne, R.W.; Glauert, Audrey M.; Dingle, J. T.; Lucy, J. A.

doi:10.1038/196952a0

Cited by 420 publications

(196 citation statements)

References 7 publications

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“…For the 5 or 10 nm gold-streptavidin conjugates (British BioCell Intl. ), which likely exceed the size of the pores made by saponin, 45 myotubes were first fixed with 4% paraformaldehyde for 10 min, and then permeabilized with Triton X-100 (Sigma; 0.1% in PBS) for 30 min. The cells were then incubated with 5 or 10 nm goldstreptavidin (1:100 in PBS) for 2-12 h followed by incubation with biocytin-Alexa568 (Invitrogen; 1:20,000) for 30 min to fluorescently label the streptavidin-gold conjugate.…”

Section: Methodsmentioning

confidence: 99%

Bimolecular fluorescence complementation and targeted biotinylation provide insight into the topology of the skeletal muscle Ca²⁺channel β1a subunit

et al. 2012

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2+ , calciumIn skeletal muscle, L-type calcium channels (DHPRs), localized to plasma membrane sarcoplasmic reticulum junctions, are tightly packed into groups of four termed tetrads. Here, we have used bimolecular fluorescence complementation (BiFC) and targeted biotinylation to probe the structure and organization of b1a subunits associated with native Ca V 1.1 in DHPRs of myotubes. The construct YN-b1a-YC, in which the non-fluorescent fragments of YFP ("YN" corresponding to YFP residues 1-158, and "YC" corresponding to YFP residues 159-238) were fused, respectively, to the N-and C-termini of b1a, was fully functional and displayed yellow fluorescence within DHPR tetrads after expression in b1-knockout (b1KO) myotubes; this yellow fluorescence demonstrated the occurrence of BiFC of YN and YC on the b1a N-and C-termini. In these experiments, we avoided overexpression because control experiments in non-muscle cells indicated that this could result in non-specific BiFC. BiFC of YN-b1a-YC in DHPR tetrads appeared to be intramolecular between Nand C-termini of individual b1a subunits rather than between adjacent DHPRs because BiFC (1) was observed for YN-b1a-YC co-expressed with Ca V 1.2 (which does not form tetrads) and (2) was not observed after co-expression of YN-b1a-YN plus YC-b1a-YC in b1KO myotubes. Thus, b1a function is compatible with N-and C-termini being close enough together to allow BiFC. However, both termini appeared to have positional freedom and not to be closely opposed by other junctional proteins since both were accessible to gold-streptavidin conjugates. Based on these results, a model is proposed for the arrangement of b1a subunits in DHPR tetrads.

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Section: Methodsmentioning

confidence: 99%

Bimolecular fluorescence complementation and targeted biotinylation provide insight into the topology of the skeletal muscle Ca²⁺channel β1a subunit

et al. 2012

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“…The incorporation of saponins into cell or endosomal membranes might expose the incorporated antigen to cytosolic proteases. According to Bangham et al(1962), saponins have been shown to intercalate into cell membranes, through interaction with cholesterol, forming 'holes' or pores. It is currently unknown if the adjuvant effect of saponins is related to pore formation, which may allow antigens to gain access to the endogenous pathway of antigens presentation, promoting cytotoxic T-lymphocyte (CTL) response (Sjölander et al, 2001).…”

Section: Mechanism Of Actionmentioning

confidence: 99%

Adjuvant effects of saponins on animal immune responses

Rajput

Xiao

et al. 2007

J. Zhejiang Univ. - Sci. B

223

149

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Abstract:Vaccines require optimal adjuvants including immunopotentiator and delivery systems to offer long term protection from infectious diseases in animals and man. Initially it was believed that adjuvants are responsible for promoting strong and sustainable antibody responses. Now it has been shown that adjuvants influence the isotype and avidity of antibody and also affect the properties of cell-mediated immunity. Mostly oil emulsions, lipopolysaccharides, polymers, saponins, liposomes, cytokines, ISCOMs (immunostimulating complexes), Freund's complete adjuvant, Freund's incomplete adjuvant, alums, bacterial toxins etc., are common adjuvants under investigation. Saponin based adjuvants have the ability to stimulate the cell mediated immune system as well as to enhance antibody production and have the advantage that only a low dose is needed for adjuvant activity. In the present study the importance of adjuvants, their role and the effect of saponin in immune system is reviewed.

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“…The toxic effects of saponins are due to their ability to form complexes with membrane sterols (Bangham and Horne, 1962). In ruminants, they inhibit rumen fermentation and may cause ruminant bloat (Sen et al, 1998), apparently through the inhibition of some rumen bacteria and protozoa (Wang et al, 2000;Wina et al, 2006).…”

Section: Introductionmentioning

confidence: 99%

Bacterial detoxification of saponins in the crop of the avian foregut fermenter <i>Opisthocomus hoazin</i>

García-Amado¹,

Michelangeli²,

Gueneau³

et al. 2007

J. Anim. Feed Sci.

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The hoatzin is a folivorous bird with microbial fermentation in the crop that consumes plants that contain saponins. We investigated detoxifi cation activity of saponins by bacteria from the hoatzin crop, and how this activity was affected in the presence of methanogens. Strains and mixed cultures were grown in presence of 200 μg·ml -1 Quillaja saponins. Detoxifi cation was determined as loss of haemolytic activity. The methanogenesis was inhibited added bromoethanesulphonic acid (BES). Mixed cultures showed higher rate of saponin detoxifi cation than strains and it was reduced by BES. The hoatzin bacteria detoxify saponins, and methanogenic Archaea might act as a hydrogen sink.

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Action of Saponin on Biological Cell Membranes

Cited by 420 publications

References 7 publications

Bimolecular fluorescence complementation and targeted biotinylation provide insight into the topology of the skeletal muscle Ca²⁺channel β1a subunit

Bimolecular fluorescence complementation and targeted biotinylation provide insight into the topology of the skeletal muscle Ca²⁺channel β1a subunit

Adjuvant effects of saponins on animal immune responses

Bacterial detoxification of saponins in the crop of the avian foregut fermenter <i>Opisthocomus hoazin</i>

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Action of Saponin on Biological Cell Membranes

Cited by 420 publications

References 7 publications

Bimolecular fluorescence complementation and targeted biotinylation provide insight into the topology of the skeletal muscle Ca2+channel β1a subunit

Bimolecular fluorescence complementation and targeted biotinylation provide insight into the topology of the skeletal muscle Ca2+channel β1a subunit

Adjuvant effects of saponins on animal immune responses

Bacterial detoxification of saponins in the crop of the avian foregut fermenter <i>Opisthocomus hoazin</i>

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Product

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Bimolecular fluorescence complementation and targeted biotinylation provide insight into the topology of the skeletal muscle Ca²⁺channel β1a subunit

Bimolecular fluorescence complementation and targeted biotinylation provide insight into the topology of the skeletal muscle Ca²⁺channel β1a subunit