Halobacterium

Animals cannot synthesize vitamin A-active compounds and necessary quantities are obtained by ingestion of vitamin A or by consumption of appropriate provitamin A compounds such as P-carotene. Carotenoids are manufactured exclusively by plants and photosynthetic bacteria. Until the discovery of vitamin A in the purple bacterium Halobacterium halobium in the 1970s, vitamin A was thought to be confined to only the animal kingdom (56). Table 4 Hsts RDA and U.S. RDA for vitamin A (67). 

ITowever, membrane proteins can also be distributed in nonrandom ways across the surface of a membrane. This can occur for several reasons. Some proteins must interact intimately with certain other proteins, forming multisubunit complexes that perform specific functions in the membrane. A few integral membrane proteins are known to self-associate in the membrane, forming large multimeric clusters. Bacteriorhodopsin, a light-driven proton pump protein, forms such clusters, known as purple patches, in the membranes of Halobacterium halobium (Eigure 9.9). The bacteriorhodopsin protein in these purple patches forms highly ordered, two-dimensional crystals. 

FIGURE 10.20 A schematic drawing of Halobacterium halobium. The purple patches ( tain bacteriorhodopsin (bR). 

If a phylogenetic comparison is made of the 16S-Iike rRNAs from an archae-bacterium Halobacterium volcanii), a eubacterium E. coli), and a eukaryote (the yeast Saccharomyces cerevisiae), a striking similarity in secondary structure emerges (Figure 12.40). Remarkably, these secondary structures are similar despite the fact that the nucleotide sequences of these rRNAs themselves exhibit a low degree of similarity. Apparently, evolution is acting at the level of rRNA secondary structure, not rRNA nucleotide sequence. Similar conserved folding patterns are seen for the 23S-Iike and 5S-Iike rRNAs that reside in the... 

Even entrapment of entire cells within reversed micelles without loss of their functionality has been achieved. For example, mitochondria and bacteria (Actinobacter cal-coaceticus, Escherichia coli, Corynebacterium equi) have been successfully solubilized in a microemulsion consisting of isopropyl pahnitate, polyoxyethylene sorbitan trioleate [162], Enhanced hydrogen photoproduction by the bacterium Rhodopseudomonas sphaeroides or by the coupled system Halobacterium halobium and chloroplasts organelles entrapped inside the aqueous core of reversed micelles with respect to the same cells suspended in normal aqueous medium has been reported [183,184],... 

Phospholipids in purple membrane of Halobacterium Silica gel G halobium Note H = hexane, D = diethyl ether, F = formic acid, BuOH = butanol. 23... 

Ethanol and choline glycerolipids were isolated from calf brain and beef heart lipids by PTLC using silica gel H plates. Pure ethanol amine and choline plasmalogens were obtained with a yield of 80% [74]. Four phosphohpid components in the purple membrane (Bacteriorhodopsin) of Halobacterium halobium were isolated and identified by PTLC. Separated phosphohpids were add-hydrolyzed and further analyzed by GC. Silica gel G pates were used to fractionate alkylglycerol according to the number of carbon atoms in the aliphatic moiety [24]. Sterol esters, wax esters, free sterols, and polar lipids in dogskin hpids were separated by PTLC. The fatty acid composition of each group was determined by GC. 

Henderson, R., The purple membrane from Halobacterium halobium, Ann. Rev. Biophys. Bioeng., 6, 87 (1977). 

Kushwaha, S. C., J. K. G. Kramer, and M. Kates. 1975. Isolation and characterization of C50-carotenoidpigments and other polar isoprenoids from Halobacterium cutirubrum. Biochim. Biophys. Acta. 398 303-314. 

Indeed, hydrophilic N- or C-terminal ends and loop domains of these membrane proteins exposed to aqueous phases are able to undergo rapid or intermediate motional fluctuations, respectively, as shown in the 3D pictures of transmembrane (TM) moieties of bacteriorhodopsin (bR) as a typical membrane protein in the purple membrane (PM) of Halobacterium salinarum.176 178 Structural information about protein surfaces, including the interhelical loops and N- and C-terminal ends, is completely missing from X-ray data. It is also conceivable that such pictures should be further modified, when membrane proteins in biologically active states are not always present as oligomers such as dimer or trimer as in 2D or 3D crystals but as monomers in lipid bilayers. 

The CP MAS NMR spectroscopy has been also extensively used for studies of proteins containing retinylidene chromophore like proteorhodopsin or bacteriorhodopsin. Bacteriorhodopsin is a protein component of purple membrane of Halobacterium salinarium.71 7 This protein contains 248 amino acids residues, forming a 7-helix bundle and a retinal chromophore covalently bound to Lys-216 via a Schiff base linkage. It is a light-driven proton pump that translocates protons from the inside to the outside of the cell. After photoisomerization of retinal, the reaction cycle is described by several intermediate states (J, K, L, M, N, O). Between L and M intermediate states, a proton transfer takes place from the protonated Schiff base to the anionic Asp85 at the central part of the protein. In the M and/or N intermediate states, the global conformational changes of the protein backbone take place. 

Zeth, K., Offermann, S., Essen, L.-O., and Oesterhelt, D. 2004. Iron-oxo clusters biomineralizing on protein surfaces structural analysis of Halobacterium salinarum DpsA in its low- and high-iron states. Proceedings of the National Academy of Sciences of the USA 101 13780-13785. 

In this paper, we will describe one of examples, where artificial archaeal glycolipids are applied to the construction of nano-devices containing energy-conversion membrane proteins, by employing the phytanyl-chained glycolipid we have recently developed, i.e., l,3-di-o-phytanyl-2-o- ((3-D-maltotriosyl) glycerol (Mab (Phyt)2, Fig. 1) [16,17] and natural proton pump, bacteriorhodopsin (BR) derived from purple membranes of the extremely halophilic archaeon Halobacterium salinarium S9 [18],... 

Figure 5. Chemical structures of main lipids of purple membranes from Halobacterium salinarium S9 phosphatidylglycerophosphate (PGP), phosphatidylglycerol (PG) and glycolipid sulfate (GLS).

The first transporter of this type characterised as an iron-supply system that functions in the absence of any siderophore was the Sfu system of S. marcescens [224]. Later, similar systems were reported from Neisseria gonorrhoea and Neisseria meningitidis, and have been detected by analysing the genomes of a variety of bacteria, e.g. Actinobacillus pleuropneumoniae, B. halodurans, Campylobacter jejuni, Ehrlichia chaffeensis, Halobacterium sp., H. influenzae,... 

Halobacterium salinarum Rapid expression Colorimetric expression Easy scale-up Cloning and transformation complicated Requires fusion protein strategy Lack of post-translational modifications... 

Halobacterium salinarum Easy Low/easy Low Short/easy High Low... 

Halobacterium Requires high salt concentrations for growth... 

---