Valinomycin, structure

The transmembrane potential derived from a concentration gradient is calculable by means of the Nemst equation. If K+ were the only permeable ion then the membrane potential would be given by Eq. 1. With an ion activity (concentration) gradient for K+ of 10 1 from one side to the other of the membrane at 20 °C, the membrane potential that develops on addition of Valinomycin approaches a limiting value of 58 mV87). This is what is calculated from Eq. 1 and indicates that cation over anion selectivity is essentially total. As the conformation of Valinomycin in nonpolar solvents in the absence of cation is similar to that of the cation complex 105), it is quite understandable that anions have no location for interaction. One could with the Valinomycin structure construct a conformation in which a polar core were formed with six peptide N—H moieties directed inward in place of the C—O moieties but... 

Valinomycin is a macrocyclic dodecadepsipeptide with 12 subunits — amino- and hydroxycarboxylic acids — which are connected by alternate peptide and ester bonds. It consists of three identical fragments D-Hylv-D-Val-L-Lac-L-Val (Fig. 1). Although the valinomycin structure has been discussed in a recent review on complex formation of monovalent cations with biofunctional ligands it is described here in some detail... 

The X-ray crystal structures of many of these complexes have now been determined representative examples are. shown in Fig. 4.11 from which it is clear that, at least for the larger cations, coordinative saturation and bond rhrectionality are far less significant factors than in many transition element complexes. Further interest in these ligands stems from their use in biochemical modelling since they sometimes mimic the behaviour of naturally occurring, neutral, macrocydic antibiotics such as valinomycin, monactin, nonactin, nigericin... 

With respect to the carrier mechanism, the phenomenology of the carrier transport of ions is discussed in terms of the criteria and kinetic scheme for the carrier mechanism the molecular structure of the Valinomycin-potassium ion complex is considered in terms of the polar core wherein the ion resides and comparison is made to the Enniatin B complexation of ions it is seen again that anion vs cation selectivity is the result of chemical structure and conformation lipid proximity and polar component of the polar core are discussed relative to monovalent vs multivalent cation selectivity and the dramatic monovalent cation selectivity of Valinomycin is demonstrated to be the result of the conformational energetics of forming polar cores of sizes suitable for different sized monovalent cations. 

In what follows, the phenomenology of carrier transport will be briefly reviewed along with the mechanism of the Valinomycin model of carrier transport. The development of the molecular structure of Valinomycin will be considered in some detail, since the key to the dramatic selectivity of Valinomycin is thought to reside in the energetics of the molecular structure. Confidence in an understanding of the molecular structure of the Valinomycin-cation complex becomes tantamount to confidence in the presented basis of ion selectivity. 

Fig. 19. Space filling models of the molecular structure of the Valinomycin-potassium ion complex as originally determined.

Fig. 20. A. Conformation of the Valinomycin-cation complex derived for solution using a combination of proton magnetic resonance data and conformational energy calculations. This structure agrees within tenths of an Angstrom with the crystal structure subsequently determined (100) and shown in Fig. 21. Reproduced with permission from Ref.99).

Fig. 21. Crystal structure of the Valinomycin-K+ complex. Reproduced with permission from Ref.100). This crystal structure confirmed within tenths of an Angstrom the structure derived previously in solution 97 98) and by means of conformational energy calculations...

Fig. 23. Space filling model of the Enniatin B—K + complex after the crystal structure 103). Since the carbonyl moieties coordinating the cation are similar for Enniatin B and Valinomycin, the difference in selectivities must arise due to the energetics of the conformations required to achieve coordination of the cation...

Fig. 24. Calculation of the conformational energy of Valinomycin as a function of the size of the polar core which contains the ion. This uses the structure of Fig. 20. The verticle lines are the optimal core sizes for the indicated ions. Based on the conformational energy component, selectivity for K+ and Rb+ would be similar and Cs+ less favored. Na+ is off the curve suggesting that this conformation cannot form a polar core small enough to complex Na+ by means of this conformation. Adapted with permission from Ref.

Especially sensitive and selective potassium and some other ion-selective electrodes employ special complexing agents in their membranes, termed ionophores (discussed in detail on page 445). These substances, which often have cyclic structures, bind alkali metal ions and some other cations in complexes with widely varying stability constants. The membrane of an ion-selective electrode contains the salt of the determined cation with a hydrophobic anion (usually tetraphenylborate) and excess ionophore, so that the cation is mostly bound in the complex in the membrane. It can readily be demonstrated that the membrane potential obeys Eq. (6.3.3). In the presence of interferents, the selectivity coefficient is given approximately by the ratio of the stability constants of the complexes of the two ions with the ionophore. For the determination of potassium ions in the presence of interfering sodium ions, where the ionophore is the cyclic depsipeptide, valinomycin, the selectivity coefficient is Na+ 10"4, so that this electrode can be used to determine potassium ions in the presence of a 104-fold excess of sodium ions. 

These substances include primarily depsipeptides (compounds whose structural units consist of alternating amino acid and ar-hydroxy acid units). Their best-known representative is the cyclic antibiotic, valinomycin, with a 36-membered ring [L-Lac-L-Val-D-Hy-i-Valac-D-Val]3, which was isolated from a culture of the microorganism, Streptomyces fulvissimus. Figure 6.13 depicts the structure of free valinomycin and its complex with a potassium ion, the most important of the coordination compounds of valinomycin. 

Attachment of carbonyl groups to crowns makes these products more akin structurally to the natural ionophore antibiotics such as valinomycin. The dioxo-derivative (179) of 18-crown-6 was prepared in 35% yield by condensation of tetraethylene glycol and diglycolic acid chloride in benzene at 50 °C for 48 hours (Izatt et al., 1977a and 1977b). This product gives binding constants for Na+, K+ and Ba2+ in methanol which are 102—104 times less stable than for the parent crown - the lower constants are a reflection of less favourable AH values for complexation in these... 

A basic property of an ionophore is that it is capable of forming a structure with a lipophilic exterior and polar cavity, as depicted in the scheme of the structure of valinomycin in fig. 7.4. The ionophore cavity must contain less than 12 and preferably 5-8 polar groups. The final complex structure must be relatively stable, which can be attained by strengthening with hydrogen bonds. It should not, however, be too rigid if ion exchange is to be sufficiently rapid [153, 193]. 

In reply to Professor Ubbelohde, research on ionophores reported in Professor Simon s paper emerged from studies of the properties of natural compounds such as valinomycin and macrotetrolides. That confirms once more the importance of such studies as initial points for the investigations of the multidimensional world of biology. I would like to add also that the natural ionophores resemble in several respects the much larger enzymes (I) they are specific and stereospecific, (2) their active sites are located in cavities, (3) their stable secondary structure, which is essential for activity, is determined by the primary structure, and (4) they transport hydrophilic substrates through lipophilic membranes. 

The results (Table 10) show that the cryptands could act to produce carrier-mediated facilitated diffusion and there was no transport in the absence of the carrier. The rate of transport depended upon the cation and carrier, and the transport selectivity differed widely. The rates were not proportional to complex stability. There was an optimal stability of the cryptate complex for efficient transport, logKs 5, and this value is similar to that for valinomycin (4.9 in methanol). [3.2.2] and [3.3.3] showed the same complexation selectivity for Na+ and K+ but opposing transport selectivities. The structural modification from [2.2.2] to [2.2.C8] led to an enhanced carriage of both Na+ and K+ but K+ was selected over Na+. The modification changes an ion receptor into an ion carrier, and indicates that median range stability constants are required for transport. Similar, but less decisive, results have been found in experiments using open-chain ligands and crown ethers.498... 

An extensive range of synthetic analogues of valinomycin and the enniatins has been prepared and investigated these studies have been fully described in ai monograph.532 Synthetic peptides have also been prepared and their complexing properties investigated for example [L-Val-D-Pro-D-Val-L-Pro]3 reacts with potassium picrate to give an isolable 1 1 complex which is expected to have a structure similar to that of the K+ complex of (137).557 More detailed studies have been made on the complexation properties of the small synthetic peptides [L-Pro-Gly]3 and [L-Pro-Gly]4. 

Beta structures are found in many small peptides. Tire hormone oxytocin (Fig. 2-4), the antibiotics gramicidin S (Fig. 2-4) and valinomycin (Fig. 8-22), and the mushroom peptide antamanide (Box 28-B) are among these. The cyclic structures of these compounds favor formation of antiparallel (3 strands with sharp turns at the ends. Polypeptide antibiotics that have alternating... 

Because the stability constant of its complex with potassium is much greater than that with sodium, valinomycin is a relatively specific potassium ionophore. In contrast, the mushroom peptide antamanide has a binding cavity of a different geometry and shows a strong preference for sodium ions.388,390 The structure of the Na+-antamanide complex is also shown in Fig. 8-22B. The Streptomyces polyether antibiotic monensin (Fig. 8-22D),389,391 a popular additive to animal feeds, is also an ionophore. However, its mode of action, which involves disruption of Golgi functions, is uncertain 392... 

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