Tritons solubilization

Fig. 10. Absorbance-change transients at 820 nm in PS-1 particles containing PMSHj(PMS plus dithionite) induced by a series of four saturating dye-laser flashes. The redox states of the PS-1 reaction center and the reactions accompanying the series of laser flash-induced absorbance changes are illustrated at right. See text for further details. Figure source. Sauer, Mathis, Acker and van Best (1978) Electron acceptors associated with P-700 in triton solubilized photosystem I particles from spinach chloroplasts. Biochim Biophys Acta 503 127.

K Sauer, P Mathis, S Acker and J van Best (1978) Electron acceptors associated with P-700 in Triton-solubilized photosystem I particles from spinach chloroplasts. Biochim Biophys Acta 503 120-134... 

The preparation of photosystem II reaction centre complexes was carried out as described in Ref. 6 with the variations mentioned in (7) and with slight modifications (20 mg Triton per mg chlorophyll (Chi), 2.5 mg/ml Chi during Triton solubilization) aimed to the use of pea instead of spinach as starting material. 

Highly active Triton-solubilized spinach PSII membranes (BBY type, 400 iM 02/h mgChl) and reaction center core complexes extracted using B-octylglycojmanoside ... 

The reported experiments seem to indicate that the membrane integrity may be responsible for the manifestation of the effect of the lipid composition on the modification of the value of n. By dialysis of the 100,000 g supernatant of the Triton solubilized membrane against a phosphate buffer in the presence of MgCl2 and CaCl2, a reaggregation material was obtained, which could be pelleted by centrifugation at 30,000 g, resuspended in sodium phosphate and assayed for acetylcholinesterase (Morero et al., 1973). 

For Triton-solubilized plasma membranes, where both right-side out and inside-out iT-ATPase can be measured, there was also no difference in loss of iT-ATPase activity between the two types of initiation systems (Figure 8). These data together with the previous data from intact vesicles, suggested that iT-ATPase activity was lost primarily through the action of lipid radicals. 

Triton-solubilized, affintiy-purified HL still expresses glycoside cluster effect, albeit to a much lesser extent than the HL on the hepatocyte surface. The best di-valent oligosaccharide, PENTA-2,4, had 20-fold higher affinity than the mono-antennary structure, as compared to 1000-fold for the hepatocyte surface receptors, while the best triantennary oligosaccharide, NONA I, had 700-fold higher affinity over the monovalent structure, as compared to 100,000-fold enhacement for the hepatocyte system. Presumably organization of subunits in the soluble form is somewhat relaxed compared to the cell surface HLs. 

Chemical lysis, or solubilization of the cell wall, is typically carried out using detergents such as Triton X-100, or the chaotropes urea, and guanidine hydrochloride. This approach does have the disadvantage that it can lead to some denaturation or degradation of the produci. While favored for laboratory cell disruption, these methods are not typically used at the larger scales. Enzymatic destruction of the cell walls is also possible, and as more economical routes to the development of appropriate enzymes are developed, this approach could find industrial application. Again, the removal of these additives is an issue. 

Among them, 38a and 38b are water soluble and they can be considered as the non-micellar counterparts. All other ligands are water insoluble but can be solubilized in CTAB or Triton X-100 micelles. 

Huot et al. [38] used affinity chromatography to identify and partially purify an amiloride-binding protein with characteristics of the renal brush border Na /H exchanger. The high-affinity amiloride analog A35 (5-A-(3-aminophenyl)amiloride) was coupled to Sepharose CL-4B through a triglycine spacer. Rabbit renal brush border membranes were solubilized with 0.6% Triton X-100, incubated with the... 

In a preliminary report, Ross et al. [40] used affinity chromatography to identify a putative bovine renal brush border Na /H exchanger. Brush border membranes were solubilized with Triton X-100 and chromatographed sequentially over lentil lectin Sepharose 4B and 5-(A-benzyl-iV-ethyl)amiloride coupled to epoxy-activated Sepharose 6B. The eluant contained 178- and 146-kDa proteins that were susceptible to Endo-F. Moreover, the eluants reacted on dot blot immunoassays with antisera to a 20-amino acid peptide of a human Na /H exchanger vide infra). The relationship between these proteins and the 66-kDa protein previously identified by the same investigators using amiloride photolabeling is presently unclear. 

Anionic surfactants (SDS and deoxycholate) are more effective solubilizing agents at higher concentration than nonionic surfactants (Lubrol, Triton)... 

Triton X-114-Mediated Solubilization o/Vicia Faba Chloroplast Polyphenol Oxidase... 

Polyphenol oxidase occurs within certain mammalian tissues as well as both lower (46,47) and higher (48-55) plants. In mammalian systems, the enzyme as tyrosinase (56) plays a significant role in melanin synthesis. The PPO complex of higher plants consists of a cresolase, a cate-cholase and a laccase. These copper metalloproteins catalyze the one and two electron oxidations of phenols to quinones at the expense of 02. Polyphenol oxidase also occurs in certain fungi where it is involved in the metabolism of certain tree-synthesized phenolic compounds that have been implicated in disease resistance, wound healing, and anti-nutrative modification of plant proteins to discourage herbivory (53,55). This protocol presents the Triton X-114-mediated solubilization of Vida faba chloroplast polyphenol oxidase as performed by Hutcheson and Buchanan (57). 

The reader may consult Sanchez-Ferrer et al. (58), Nunez-Delicado et al. (59), and Sajo et al. (29) for updates concerning the use of Triton surfactants to solubilize membrane-bound polyphenol oxidase. In addition, Kieselbach et al. (60) offers a contemporary view regarding the isolation and characterization of chloroplast thylakoids. Wilhelnova (61) compared the abilities of different detergents to fragment thylakoid membranes. 

Triton X-100-Promoted Solubilization o/Postia Placenta Hyphal Sheath Oxalate Decarboxylase... 

Fig. 3. Flowchart showing how to use Triton-X to solubilize hyphal ODC. From Micales (65), with permission.

I didn t use the triton X-100 example because it is a neutral entity. It is fine for an organic chemist, but inorganic entities can t interact with this micelle. However, a triton X-100 micelle is very well studied because biochemists are interested in using it as a model membrane since it makes big flat-disk micelles. Biochemists consider it as resembling a section of membrane, which can solubilize proteins. It is a good model for biological systems. 

The spike protein complex of the alphaviruses appears to be assembled in the ER. Cross-linking studies of Triton X-lOO-solubilized viral proteins have shown that the p62 and El proteins are linked together to form a complex in the ER (Ziemiecki et al.. 1980). The topology of these complexes is the reverse of that in the viral particle the spikes are within the lumen of the ER and the internal domain of E2 is on the cytoplasmic side of the ER membrane. 

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