Permeases bacterial

Furthermore, if the antibiotic passes membranes through a specific port of entry, its mutational loss leads to resistance. The lack of the outer membrane protein OprD in P. aeruginosa causes resistance to the (3-lactam antibiotic imipenem. Fosfomycin passes the cytoplasmic membrane via an L-a-glycerol phosphate permease. This transport system is not essential for bacterial growth and therefore mutants with a reduced expression are frequently selected under therapy. 

The lactose transporter (lac permease) of E. coli is without doubt the most intensively studied and best understood of the bacterial proton-linked sugar transporters. Since its sequence was reported in 1980 [233] prodigious efforts have been made to elucidate its molecular mechanism by site-directed mutagenesis and other means. These studies have recently been reviewed elsewhere [234,235] and so will not be discussed in detail here. The important question for the present Chapter is whether the protein is related to the sugar-transporter family and so has lessons to teach us about their mechanisms. The permease is a 417-residue protein, and, like the other... 

When the new term permease was coined to designate bacterial membrane proteins specialized in the transport of specific metabolites [1,2], it covered a concept which was not quite new. The existence of membrane transport systems had been demonstrated in animal tissues by Cori as early as 1925 (see [3]). However, the discovery and characterization of permeases in bacteria revolutionized prospects for studying the properties of transport systems, opening the way to a new field and a very fruitful methodology. 

Ovchinnikov 234 237) has shown that bovine rhodopsin, although quite different in amino acid sequence (348 residues), also forms seven transmembrane helices. This structural similarity between bacterial and mammalian light activated membrane proteins is remarkable. Since the two amino acid sequences have little in common it would appear that the necessary requirement is seven transmembrane helices to form a channel which is specific for proton migration. For example it has been suggested that a similar arrangement and function is performed by the lactose permease of E. coli237). 

Saurin, W., Koster, W. and Dassa, E. (1994). Bacterial binding protein-dependent permeases characterization of distinctive signatures for functionally related integral cytoplasmic membrane proteins, Mol. Microbiol., 12, 993-1004. 

Likewise, for zinc, bacteria have developed active uptake systems (Hantke, 2001). In many bacteria the high-affinity Zn2+ uptake system uses an ABC transporter of the cluster 9 family, which mostly transports zinc and manganese and is found in nearly all bacterial species. First identified in cyanobacteria and pathogenic streptococci, but also found in E. coli, the system is encoded by three genes ZnuABC and consists of an outer membrane permease ZnuB, a periplasmic-binding protein ZnuA and a cytoplasmic ATPase ZnuC. Low-affinity transporters of the ZIP family, described later in this chapter, such as ZupT, have also been shown to be involved in bacterial zinc uptake. 

Two are hydrophobic intrinsic membrane domains, each with six membrane-spanning helices and two are peripheral membrane ATP binding domains. All four domains may be in a single peptide chain, as in CFTR, or they may be separate smaller proteins as in bacterial periplasmic permeases.431 446... 

Figure 2. Proposed model for a bacterial permease system...

The bacterial lactose-transport protein (lactose permease) transports j3-galactosides, such as lactose, o-nitrophenyl-jS-galactoside, and isopropyl-jS-thiogalactoside. It does not transport galactosides with an a-glycosidic linkage. 

Sakamoto, T., Inoue-Sakamoto, K., and Bryant, D. (1999). A novel nitrate/nitrite permease in the marine cyanobacterium Synechococcus sp. strain PCC. (7002). J. Bacterial. 181, 7363—7372. 

Relatively few membrane transport proteins have been structurally characterized. Some of the best understood examples to date are the lactose permease and glycerol-3-phosphate transporter and the Ca + P-type ATPase (which is a primary ion pump). Other structurally well-characterized transport proteins include the bacterial porins and siderophore receptor proteins. In addition structures have been determined for several ion channels and additional bacterial transporters that are either directly relevant to or models for proteins important in drug transport. The following web sites maintained by Hartmut Michel and Stephen White respectively, contain exceptionally useful listings of these and other solved membrane protein structures and are frequently updated ... 

Figure 13.11. Action of Lactose Permease. Lactose permease pumps lactose into bacterial cells by drawing on the proton-motive force. The binding sites evert when a lactose molecule (L) and a proton (H+) are bound to external sites. After these species are released inside the cell, the binding sites again evert to complete the transport cycle. Lactose permease is an example of a symporter.

A crucial clue to the mechanism of gene regulation was the observation that two other proteins are synthesized in concert with P-galactosidase—namely, galactoside permease and thiogalactoside transacetylase. The permease is required for the transport of lactose across the bacterial cell membrane. The transacetylase is not essential for lactose metabolism but appears to play a role in the detoxification of compounds that also may be transported by the permease. Thus, the... 

Transporters for monocarboxylic acids include (1) the bacterial lactate permease LctP family, (2) eukaryotic proton-linked monocarboxylate transporter MCT family, and (3) monocarboxylate permease. These transporters contain no ATP binding cassette (ABC) motifs and are considered to transport monocarboxylic acids via a proton-coupled reaction (Nakano, Fukaya, and Horinouchi, 2006). 

Considerable progress has been made over the past 10 years by the group of Kaback at UCLA, who have taken numerous biophysical approaches toward studying the structure of the bacterial transporter lactose permease (136). Table 8.3 lists the techniques that this and other groups have used to solve the structure and topology of membrane transporters in the absence of a crystal structure. 

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