On the mechanism of saponin hemolysis—I

Segal, Ruth; Shatkovsky, Puah; Milo-Goldzweig, Ilana

doi:10.1016/0006-2952(74)90027-6

Cited by 66 publications

(17 citation statements)

References 12 publications

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“…5) Some authors have postulated that the last step consists of an enzymatic deglycosylation that releases the aglycon and produces cell lysis. 6,7) Aglycons can show weak hemolytic activity but are not solely responsible for cell destruction. While the aglycon is very important in considering structure-activity relationships, the contribution of the sugar moiety is also significant.…”

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confidence: 99%

Synthesis and Hemolytic Activity of Some Hederagenin Diglycosides

Chwalek

Plé

Voutquenne-Nazabadioko

2004

Chem. Pharm. Bull.

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Saponins are steroid or triterpenoid glycosides which are widespread in terrestrial and marine plants and possess various biological and pharmacological activities. 1) One of the first known activities of saponins is their ability to lyse erythrocytes.1) This hemolytic activity has long been studied and the origin of the phenomenon is generally ascribed to their amphiphilic properties, but the exact mechanism is not yet known especially when considering the structural diversity of saponins. It has been shown that saponins are capable of interacting with membrane cholesterol 2-4) forming pits and holes which produce a destabilisation of the membrane by a micellar type-arrangement. 5) Some authors have postulated that the last step consists of an enzymatic deglycosylation that releases the aglycon and produces cell lysis.6,7) Aglycons can show weak hemolytic activity but are not solely responsible for cell destruction. While the aglycon is very important in considering structure-activity relationships, the contribution of the sugar moiety is also significant. 8) Hemolytic activity depends on the nature, number and sequence of the sugars in the saponin.Our laboratory has long been interested in saponin isolation and identification, and, more recently, in their hemolytic structure-activity relationships. The study of hemolytic activity is a way of gaining insight into how saponins react at the membrane surface. We have compared many saponins with various triterpenoid aglycons and, as Schlösser and Wulff,8) have established that a polar substitution in ring A and weak polar substitution in rings D and E increases hemolytic activity and that monodesmosidic triterpenoid saponins are generally more active than bidesmosidic ones. 9)Structure-activity relationships of some synthetic saponins of oleanolic acid, 10-12) methyl oleanolate 13) and of other triterpenoid or steroid aglycons 14,15) have been previously established. It is thus known that the effect of the sugar residue on hemolytic activity is not transferable from one aglycon to one another. We wished to complete our study by analyzing the influence of the sugar units on hemolytic activity using hederagenin as the exclusive aglycon. Hederagenin saponins are largely represented in nature and possess many biological activities such as hemolytic, antiviral, antimicrobial, fongicidal, molluscicidal, or cytotoxic.1) With aglycons such as hederagenin, possessing a carboxylic group in position 28, it is known that hemolytic activity is greater when the free acid is esterified as a methyl ester. Unfortunately, no comparative structure-activity relationship have been performed on both the free acid and the methyl ester saponins to establish if the same factors systematically influence hemolytic activity. 8,16) We thus wished to synthesize the natural saponin 3hederagenin (isomaltosyl-hederagenin) 6 (Chart 1), and their corresponding methyl esters (1a, 2a, 3a, 4a, 5a, 6a). Compound 1 has been isolated from Fatsia japonica 17) and from the leaves of Barbarea vulgaris and ...

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confidence: 99%

Synthesis and Hemolytic Activity of Some Hederagenin Diglycosides

Chwalek

Plé

Voutquenne-Nazabadioko

2004

Chem. Pharm. Bull.

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“…If the OM of S. hyodysenteriae contained significant levels of cholesterol, as suggested by the extraction data, this membrane should be sensitive to agents that interact directly with sterols, such as the saponin digitonin. These compounds form a complex with cholesterol (20) and disrupt erythrocyte membranes (27,28). To see what effect digitonin had on S. hyodysenteriae, cells were incubated with various concentrations of digitonin.…”

Section: Resultsmentioning

confidence: 99%

Purified outer membranes of Serpulina hyodysenteriae contain cholesterol

et al. 1997

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We have isolated outer and inner membranes of Serpulina hyodysenteriae by using discontinuous sucrose density gradients. The outer and inner membrane fractions contained less than 1 and 2%, respectively, of the total NADH oxidase activity (soluble marker) in the cell lysate. Various membrane markers including lipooligosaccharide (LOS), the 16-kDa outer membrane lipoprotein (SmpA), and the C subunit of the F 1 F 0 ATPase indicated that the lowest-density membrane fraction contained outer membranes while the high-density membrane fraction contained inner membranes and that both are essentially free of contamination by the periplasmic flagella, a major contaminant of membranes isolated by other techniques. The outer membrane fractions ( ‫؍‬ 1.10 g/cm 3 ) contained 0. Swine dysentery, caused by Serpulina (formerly Treponema) hyodysenteriae (9, 10), is a highly infectious, acute to chronic disease that can result in a debilitating mucohemorrhagic diarrhea and death. The first stage of the disease is spirochetal colonization and proliferation in the enteric epithelium, resulting in a significant decrease in the absorption of nutrients, ions, and water (12). However, the spirochetes do not invade beyond the lamina propria, and putative virulence factors, such as hemolysins and lipooligosaccharide (LOS), probably cause the cellular damage (16,(21)(22)(23)(24)35). Because a close association of the bacteria with the intestinal lining is critical in the development of disease, surface components of the spirochetes must play a pivotal role in the process.Previously, different groups have attempted to isolate outer membranes (OMs) from S. hyodysenteriae with less than favorable results. This was mainly due to the contamination of OM fractions by periplasmic flagella. For example, Joens et al. (11) found seven major proteins, ranging from 42 to 32 kDa in size, in OMs isolated from S. hyodysenteriae B204 by a Sarkosyl extraction technique. However, six of the seven bands appeared to be derived from the periplasmic flagella. Similarly, Chatfield et al. (5) and Wannemuehler et al. (39) found that the majority of the putative OM proteins isolated with detergent ranged in size from 45 to 29 kDa and strongly resembled the protein profile observed for the flagellar proteins. These data suggested that membranes isolated by these techniques were heavily contaminated by flagella. Therefore, a more effective technique is needed to better define the OM of S. hyodysenteriae.We have developed a membrane separation technique that permits the isolation of S. hyodysenteriae OMs and inner membranes (IMs) with minimal contamination by flagella. Analysis of the OMs indicated that they are quite interesting. First, the OM of S. hyodysenteriae has a lower density than the IM, which is very unusual for bacteria. In addition, the OM contains significant levels of cholesterol. Previously, Mycoplasma species (6), Borrelia hermsii (15), and S. hyodysenteriae (30, 33) had been described as having sterols in their membranes; however, the distributions ...

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“…In addition, there is electron microscopic evidence that saponintreated red cells have folded cell membranes and form pits as well as vesicles [5,8]. If the first phase (Stl) of the hemolytic process is the period of absorption of saponin to the red cell membrane, as stated by Segal et al [17], the prolonged lag phase after storage might suggest a loss of affinity between saponin and the membrane substance which has a protec tive action against osmotic stress. Reed and co-workers [2,11] reported a progressive de crease of the total lipid, phospholipid and cholesterol per cell during the first 21 days of storage.…”

Section: Hypotonic Hemolysis Of Storage ---------------------------mentioning

confidence: 99%

“…During treatment with saponin, a lag time precedes swelling and hemolysis. Segal et al [17][18][19] suggested that saponin hemolysis was an enzyme-me This report describes the changes of sa ponin hemolysis curves during blood storage and suggests that saponin hemolysis might be useful for research on red cell membrane properties.…”

Section: Introductionmentioning

confidence: 99%