ai 12 Hydrophobic and hydrophilic molecules Molecules can be divided into (wo main categories as egards their hydration: (1) those without charged groups or atoms (O, N, etc.) capable of forming hhydrogen bonds ~ these are described as hydrophabie and are mostly molecules with aliphatic and aromatic hydrocarbon chains; (2) those which, because they are charged or contain nucleophilic groups, can easily bind water molecules ~ these are hydrophilic substances, ‘The mechanisms by which the two groups become hyeirated differ markedly from cach other and need to be analysed in detail. 2 12.1. Hydration of hydrophobic groups When a hydrocarbon is transported from an organic to an aqueous phase, @ thermodynamic study shows thatthe corresponding AGis postive (unfavourable reaction). When the two phases are in equilibrium, their chemical potentials are equal. IF, isthe molar faction ofthe hydrocarbon in the organiephase ad Xwthe same in the aqueous phase, we have (sce Box 12.1: H+ RPInXy = 18, + RTI, (2.4) RTI Xu/X, Hoe (122) where and jg are the standard chemical potentials ofthe hydrocarbonin the two phases. Since AG is postive, § < jay and hence Xy < Yq, $0 that only a very small faction is dissolved in the water. I, for example, AG=4kealmot, Xq/Xo— 1.3 10° at room temperature. The positive value of AG isa result of the high value of 4S, even though A//is often negative. Note thatthe measurements of the thermodynamic quantities must take account of the way in which the solute is transported from one phase to the other, my)i 12.1 HYDRATION GF HYDROPHOBIC GROUPS 261 Box 12.1 Transition from an organic to an aqueous phase ‘The chemical potntil jy of the hydrocarbon solution should be writen inits exact form as = tn =H + RTIX + RT Inf _ where fy is the activity coefficient. ‘Because ofthe very ow solubility ofthe hydrocarbon in water. we can take Ju =1 and RTInf = 9, since the term represents that part of the chemical ‘potenti resulting from interactions ofthe solute molecules with themselves. ‘Moreover the term R7'in isa purely statistical contribution to jy arising ‘rom the entropy of mixing to components assumed to have the same size of ‘molecule, The term as therefore the only one representing thefeeeneray of {heiteracton between the dissolved molecule and water, : Similarly, forthe same hydrocarbon in an organie solvent: Ys 5+ RTInXy + RT If (123) ‘Assuming that jis almost independent ofthe typeof solvent, eqn 12,3 can still be applied to a pure liquid hydrocarbon. We then have In X~0, Inf, =0 and = a3. The difference jj14° can then be measured by the transfer of liquid hydrocarbon from the pure liquid to the aqueous solution, For n-alkanes, it is found that Ben = -2436—884n, (124) where jis in eal mol and 9, i the number of earbon atoms, is interesting to compare the values of Aye obtained for n. ~ 5 (pentane) and ‘ne=6 (hexane) with those for non-linear or substituted hydrocarbons with the same value of) ne n=6 pentane 6856 hexane 7740 eyelopentane 6000 cyclohexane 6730 methyleyelopentane 6880. methyleyelohexane —7730 Saturated eyctchydrocarbonsarelesshydrophobic than linear ones, but the simple sudition of « methyl group increases Ay by between 08 and 0.9kcal mol”! and reaches value close to that for linear hydrocarbons. ‘Another comparison can be made with aromatic hydrocarbons having =6. For benzene, Aji = 4620 and for toluene Ayi=—S430, so thot these are Iss hydrophobie than the saturated near compounds. probably becauseof the van der Waals iterations between the 7 electrons of the carbon ring and the water ‘molecules. The difference of 0.8keal mol~ between benzene and toluene reflects that found above forthe addition ofa methyl group. Inamixturewith feomponents, extensive quantities such asentropy and volume can be associated with intensive quantities known as portil molar guanies, For example, the partial molar entropy ofthe ith component is defined by262 MOLECULAR BIOPHYSICS = (-8G/OT)p,, we have that (Si) = -8/8N(06/2T) px, = ~A/9T(G/ON) py, ou /oT i + RT X,, where X;is the molar fraction ofthe ith component, we (Si) = -RInXs~(OH6/OT) py, (23) ‘The ~RIn.X; term, known as the cratic contribution, arises from the mixing entropy. The term 5} = (HOT, (126) is the unitary entropy. Applying theserelationshipsto the bimolecular reaction A + B= ABand noting that one AB molecules obtained for each pair of A and Bmofecules, we have fora molar solution (¢;= 1) AS/= AS? + RInX, (2.7) For water, ¢|= 55.6, so X;= 1/556 and hence RTIn X= Sealmol"'K-!. At 300K, this erate term contributes ~2,4 kcal mol” tothe free energy. Moreover, the change in unitary entropy is AS; Si Rin (2s) ‘This entropy change is true representation of the hydration process and isthe {quantity used in what follows. For example, for n-butane AH = ~0.85 keal mol”, AS=~223calmol'K-! and AG=S9kealmol!, More generally, for all aromatic and aliphatic hydrocarbons, 2.9 < AH < 0.7kealmol! and —23 < AS < ~18ealmol-!K~ and hence 2.6 < AG < 6keal mol”. The large negative value of AS reflects the organization of water molecules around hydrophili groups 10 form ‘cages’. In other words, the contact between water and the hydrophobic group ismuch less favoured than contact between two water molecules. Moreover, this solvation of a hydrophobic group is accompanied by a decrease in volume due toa better use of the available space. Its therefore impossible to conceive ofthe water molecules arranged around the hydrophobic group as they are in ice 1 berg model), When waters frozen at 273K, the latent heats 0a giving ~1.44keal mol! and AS = —5.3calmol~' Kt would therefore need four water molecules to be ‘frozen’ to give a value for AS of around ~20calmol-'K~'. The corresponding value of AH would then be about ~58keal, which is much too high. In the recent two-state model (tetra-coordinated A and pente-coordinated B), theintroduction of hydrophobic compound would endo inerease theamountof A (the state with the lowest energy and entropy and the largest volume). ‘With most solvents, the solute takes the place of solvent moleeules inthe liquid network, but in water a hydrophobic solute cannot replace a water molecule. Instead ofthe substitution, therefore, an interstitial compound is formed and this is ‘hy steric and spatial characteristics ofthe molecules are so important. Similarly, the“loosest' structure will be the one that accommodates the solute molecules most casily, leading tothe high negative value of AS, The favourable change in enthalpy (SH < 0) isanother way of expressing the structural rearrangement in water.i 12.2. HYDRATION OF IONS 263 12.2 Hydration of ions 12.2.1. Structural aspect In the strong eletre field (of the order of 10"° Vm") near an ion, the permanent ipole ofthe water molecule take up direction pointing towards the central ion. ‘The orientation energy IV'=—p- E. where pis the dipole moment of the water molecule is between 10k,7-and 207. Inthe first hydration shell around the ion, the water molecules ean no longer rotate freely and because of this they Form @ structure very diferent from that ince I called the first hydration shell or ordered vegion 1 (Frank and Wen, 1957). Between this ordered layer and the bul. of the ‘water there must be a second and somtewhat disordered region If, since local order similar to that revealed by X-rays cannot be established. Tons can be classified according to the relative importance of layers I and Il ‘With 2 large ratio of charge to ‘onic rads, the number of ordered water mole- culesin region lis greater than in Hand the salt solution will be more ordered than ‘pure water: this isso for Li*, F- and Mg”*. For Cs*, Rb* and C103, on the other hand, the salt solution will be more ‘liquid’ than pure water, i. it has les local order. Such a qualitative description is just as valid as the previous one based on parameters like the radius of the hydrated ion and the hydration number, whose values depend mainly on the method used. ‘Another approdcls has more recenily been proposed: the introduction ofa cor- relation sme’ between the reorientating motions of water molecules, which ean be measured by NMR. It is then possible to caleulate 7, in the hydration sphere (layer I) and to compare it with the value (rin pure water. Infact, we find that ‘re (rep for structure-making ions and 1. < (reo for structure-breaking ions. In cother words, experiment shows that 1/7) increases with concentration in the first case and decreasesin the second case, T being the longitudinal relaxation time of the proton. 12.22 Thermodynamic aspect ‘Thefiee energy ofhydration AG, for an ion can bedefinedas the work required to rove the ion from vacuum ()=1) to a medium of dieletsic constant ey Approximately equal 080, An expression for AGy has been given by Bor (see eq 1537) AGu = (¢ (8200) (1~« (129) ‘where gis the ioniccharge and the ionic radivs, Born used the following processto caleulate AG: (1) remove the charge from the ion in vacuum (work AM}: (@) transfor the uncharged ion from vacuum to water (work AV, assumed 10 be zero}, (G)_ recharge the ion in water (work AW). However, we have to decide what we mean by the ionic radius. IF we take it ‘equal 1 that determined by crystallographic measurements the caleulated values for AGare oo high compared with the experimental ones. The discrepancy can be ‘accounted for by several factors: (i) r should be increased tothe value given by its vvan der Waals radius, thus taking betieraecount of theionic hindrance in solution:264 Electron density Figure 124 Econ deny dirbuton inanakabalce ))) MOLECULAR BIOPHYSICS (ithe value of 4 W’sshould be different from zero; and (ii) ought to be reduced ‘ecause near thoion the water moleculesnolonger have the rotational freedom they have in free water thus lowering the diclectrie constant. However it appearspreferable (Rashin and Honig, 1985) to reconsider the Born process. Take, for example, the discharging of cation with charge-+e in nacwo, for ‘which an electron must be added to obtain a neutral atom. Transporting this atom into water is equivalent to.creating a spherical cavity of radius R inside which the cleetron density of the solvent is negligible. To evaluate R, we can call on the ‘electron density distribution insidean ionicerystal ofthe typeX”M>, where X~ isa halogen and M™~ an alkali (Fig. 1.1), The electron density due to a cation M* located at E becomes significant ata point D such that AD (with X-at A)isequal to the anionic radius. On the other hand, because the eletton cloud of X~ cannot penetrate the fee valence orbital of M~. the electron density of X~ becomes sig- nificant only ata distance from the centre of M* equal to the radius EB ofthis orbital. Because of thisasymmetry between X~ and M*, different radi are proposed for the cavities of anions and cations, For the cation the radius is equal to that of the “orbital, i. almost the van der Waals radius. For the anion, the radius equal 10 ‘that ofthe ion, The validity of these assumptions can be tested by comparing cal- culated and observed values ofthe hydration enthalpies ofeach ion, obtained from ‘measurements on salts Since AG = AH-TAS, AH F /Sreyr1-1/e(T/2) de 47] (1210) Hoy. vatiesasy~!, but thecaleulated values aresystematically 6-7 percent higher than those observed, leading to a similar correction factor of the same order for both eavity radii to obtain agreement between AHajc and AH. The results are listed in Table 12.1, and the experimental points now lie very close toa straight line given by AH = Ky! where Kisa constant (Fig. 12.2). We ean also consider the entropy change taking place when an ion dissolves in ‘water (change in partial molar entropy), taking the value zero for a proton. \GT WAG) AT, so that from eqn 12.9: AH ‘able 124 Corre! ove rai ancl hycaton enthabias ton Cocteau (A) ~All Tone raaus (A) ro 12 726 ast Nat 168 985 096 a ait Tea 133 Rot 2st 734 148 cs" 251 576 188, ge 145, 4624 085 186 384 98 134 492 om 151 435, ost 1st 400 at mm 142 119s 134 194 853 ta co 208 738 198 r 238 63 ent on 150 10912.2. HYDRATION OF IONS 05 06 07 08 08 “able 12.2 Untary ontop of ons eae") ire we = = 103 Nat 64 se aa 52 e 165 cath -2 na ney 0 Mae 362 a Fo zor W ° “28 ost 238 ‘Table 12.2 gives values of AS for various series of ations and anions. The sign of Sis directly related to the ‘structure-making? or ‘structure-breaking” character ofthe ions, ‘Another experimental method of studying the structural changesin liquid water produced by ions in solution isto measure the viscosity 1 of salt solutions and to compare this with the viscosity my of pure water at the sime temperature. A relationship nj = 1+ del? + Be is obtained, where cis the molar concentration of the salt, and A is a postive cosfficent reflecting the electrostatic interactions between ious that contribute 10 the energy of fluid flow. The coefficient 2, on the other hand, may be postive or negative and can be interpreted in structural terms. I, for example, Bis negative (ovhich oecurs only in the presence of eertain ions), the viscosity ofthe solution may 1) ‘il 265 Figure 122 Variaono he ryaaton ral fos win er comected ad The values ar related one ebemeniany charge (Flom Aashin ana Honig. 1985, Fepried th permison rom Jounal (oF Pysica Chamisry © 1985 American (Chamca Scien)265 Figure 123 Variavonf ho unitary Ionic ertopy Swit coetclentot viscosty MOLECULAR BIOPHYSICS Table 123 Vahes ol the coatickre B u Na K wr Fe 5 ea owe 0086 = -auor = 0007-008 aes oz & Ma o = 1 NOs 50, ness 0285 nar -onaz 0088-0046 -0200 10 on 02 as 4 be lower than that of water ata suficiently high salt concentration. Although this behaviour is dynamic rather than stati, a negative B can be interpreted as cotre- sponding tothe disappearance ofthe short-range order presentin pure water. Bean only be determined experimentally for salt solutions, i. when there is a cation: anion pair, However, a value can be attributed to each fon ia reference value chosen by assuming that K* and CI~ make equal contributions to the B of KCL solutions (Gurney, 1953) The results fora number of ions are given in Table 123, {Ifthe unitary entropy is plotted against # for each ion (Fig, 12.3), we find linear relationships defining the various families (alkalis, alkaline earths, halogens, ec.) thus implying that both parameters reflect the structural changes in liquid water. However, the distinetion between ions shows that parameters other than the clectrc field around the ion must be involved. 1223. Lyotropic series Hofmeister (1888) established a sequence of salts according to their ability co precipitate euglobulins. The salts are classified in increasing order of the con centrations needed for precipitation and hence in decreasing order of their efficiency [Nay{cittate) > Lis$Os, NaySO4, Ks POs, NasHPO, > (NH.);S0s > MgSOx > KCHsCOO, NaCHsCOO > NaCl > NaNO: my)‘i 12.2. HYDRATION OF IONS, From this itis possible to deduce an ionic order, which has since been found in ‘many other processes such as protein stabilization, enzyme inhibition and DNA stabilization, In denaturing native conformations or double-helial structures the following sequence is found, in order of increasing efficiency: + foranions SO} < CH,COO™ < Cl < Br < NOF < ClO <> < CNS” + for eations (CHs),N* < NH} < Rb*,K*.Na, Cs" < Lit < Mg < Ca < 81°" Clearly, the effet of an iom isnot related solely tits charge, and other factors must be involved. These include the ionie polarizability, which increases with size (eg. Cs" has a higher polarizability than water, the presence ofa dipole moment, ‘thepresenceof hydrophobic groupssuchas(CH,},N" andthe possibility of forming ‘additional hydrogen bonds (CIOs, NO;) with water molecules. detailed study of the local properties of water around each ofthe ions has sill tobe carried out ‘Wenow turn to phenomenon that is easier to analyse and is of great practical importance: the sating-out mechanism, We follow the treatment of Edsall and ‘Wyman (1958). The majority of water-soluble organic molecules become less soluble if water is replaced with a sat solution. Conversely, the addition of an ‘orgattic compound to salt solution lowers the solubility ofthe salt. In some eases (€g. salts of divalent cations) phase separation may occur. A denser layer rich in ‘water and salt separates from a lighter layer rich in the organic compound, his reciprocal effect on solubility has long been used forthe selective precipi- tation of proteins by the addition ofa salt such as ammonium sulphate. Itis also {involved in processes for growing biopolymer crystals, so thata detailed analysis is ‘worth undertaking. ‘Thebasicide is that theintroduction of ons into a mixture of water and organic molecules produces a rearrangement of water molecules. In the language of classical thermodynamics the adition of ions is said to reduce the activity of the \water and inerease that ofthe organie compound. The fact that charged ions are involved clearly points to the phenomenon being mainly electrical. Ina medium of diclettic constant e subjected to an electric Field E, the stored energy pet unit volume is yB/2. Theelectre field produced by the ion itself decreases by a factor Ie, Hence, ata given distance from anion of charge Ze, the energy ofthe systems lowered as the dielectric constant increases. The dieleetric constant of water (about 80) s generally higher thaa that of an organic medium, so that water molecules will bbe preferentially bound around ions tolower the energy ofthe system. This eads to an exclusion of the organic molecules (see Box 12.2. “The presence ofan organic solvent such as ethanol thus inereases the activity of the salt, and conversely a high salt concentration will increase the activity of the organiccompound and hence reduce its solubility. The reciprocity of thiseffect can casily be deduced by expressing the variation in free energy with the number of | i |i 86 pidn268 MOLECULAR BIOPHYSICS Box 12.2 Salting out ‘The derivation in this box follows Edsall and! Wyman (1958). A calculation taking all the parameters into account would be very complex. Instead, we ‘restrict the treatment to a simple model. which, although apparently unrea- listic, gives a reasonably satisfactory explanation of the observed phenomena | and allows us to predict other sslting-out mechanisms. at least qualitatively, “The medium contains n; moles of water, 1; moles ofthe organic compound and ns moles of a monovalent sal dissociated into » moles of anions and cations wit ionic charges —e and +e respectively. In a medium of dicletric constant c the electrical energy of the cation and anion are nye"/8ne=yr, and ne? Sneeor where rand rare the respectiveionic adil, assumingtheionsare spherical eiarges. The total electrical energy can be expressed as nye/Bmesoh where 1/b= Ure + Hr In water the same ion pair would havean energy given | by me’ Sazuerb where =. i the dielectric constant of water. ‘The difference between the two terms AW = (€/Sreub)(1/e-1/en)m | isthe diference between the letrcal energy ofthe on dissolved in water and dlssolved insolation. ‘The chemical potential ofthe salts the sum of @ chemical term py and an ‘lectricl term pi given by | #8, = HAW) JO; = AO/e—1ew) where A = me?/Sreph. a quantity charseterste ofthe salt ‘The chemical term is given by the classical expression in terms ofthe molar | i fraction ¥ by = 6+ RTING/¥3) | Since ns salt molecules give cations ands anions, we have X= 2n/im te + 2m) “The total chemical potential with respect to its standard valueis (Wis Hbexat = AA /2— Lew) + RT IX (X53) | In this expression, however. itis only the frst term on the right that plays any | part in the saling-out process. An electrical activity coefficient fg can be ‘defined from js’ such that = RT Inf = A(N/e-H/e~) 16 for example, we take b= 10""’m, T=300K, e=1.6% 10°C, =: | (ethanol) and 2, = 80, we find that f;-2000. The satis therefore 2000 times less soluble in ethanol than itis in water. : where the sum is overall components of the system and where iis the chemical potential (or partial molar free energy) ofthe ith component, Hence my) aqu/ 1 = 896 [Om Bnj = B/DUl 12.3 HYDRATION OF AMPHIPHILIC MOLECULES or in terms of activities, ay and a Alu /On, = Anan; ‘which gives us a direct relationship between the change in activity of one ‘component (e.g the sat) and the change in activity ofthe other (e.g. the organic compound). ‘Such an elementary treatment can only give us a general ices ofthe salting-out snechanism. In the special case of proteins, where high salt concentrations have to beused, there is no valid theory leading to-a quantitative prediction of the process. ‘Theres an empirical law of the type logs log So-KT ‘where is theionie strength, Sisthesolubility, Syitshypothetical valuefor/= and ‘Kisa constant generally proportional tothe size of the protein but also depending fon the kind of ion used. We then obtain the Hofimeister series again, as in many other phenomena. 12.3 Hydration of amphiphilic molecules 123.1. Defffition and properties of amphiphilic molecules ‘A molecule that has both a polar group and an aliphatic chain will have new sol ‘ation properties, Sucha molecule iscalled amphiphiliethe polar head wllundergo solvation like an fon, while the hydrocarhor chain will do so like a hydrophobic ‘molecule. The idea of an amphipilie molecule becomes much more important ‘whenever an interlace is created between polar and non-polar medium, An example isthe interface appearing in emulsions of eiLin water or of water i i. "The ener required to form such interfaces is generally to great forthe sytem to be sable. This s reflected by the changes occurring in the emulsion as ime passes ist a coalescence of drops ofa given medium, followed by a separation {nto two phases, as with oil and water where one phase formsa layer over the ote, depending on ther relative densities, The emulsion can be stabilized by using an “amphiphilic compound known asin emusiing agent or surfactant. By attaching, ise to each ofthe two phases the amphiphilic molecule establish a connection between the two media which reduces or eliminates the surface tension at the Interface and allows the emulsion to remain stable, possibly indefinitely. The jndusrial importance of aimphiptilic compounds can be appreciated by quoting ome igure: her annual production amounts to about | kg for every hum being on the plane. 12.32. Quatemary ammonium ions ‘Quaternary ammonium ions (NR,)*, where R isa short aliphatic chain, are good examples of compounds providing another way of studying models of hydration: by varying the length ofthe aliphatic chain and then measuring the change in ionic mobility with temperature, If short chains are bound. as in MegN~. the ionic structure-breaking effect predominates and the ionic mobility (the product of conductivity and viscosity of the solvent) must decrease with T, For jons like Bu,N* or PrN on the other hand, the cages formed around the aliphatic chains, which greatly reduce the ionie mobility at room temperature. “melt as Tinereases, thus increasing the ionic mobility (Fig. 124), If, now, an OH group is introduced ‘iii270 Figure 124 Varin of mbit wth Chemical Socey) for elongation of asc o Falende.1970.Reperea tom ey & Sons, © 1970 Jona Wey 8 Sen) temperate or quatemary ammonium bee ‘ons, (Fem Kayan Ean 888, Feprrted win permission tom Journal Figure 12.5 Varin ARP wih AS slkyammorium ons. From Lum are Biopojmes wih te permission of ohn on 08 02. SE ‘of Physical Chom @ 1988 Ameen 0 10 2 90 40 50 TEC) 148: e E Bos § -40 3 “135 ° “2-3-4 -8 AS® (eal mor") {nto the aliphatic chain, the mobility increases because partial solvation can occur without a cage effect: thus, Me(ErOF)N* has a higher mobility than MesPrN* This type of compound also gives rise to compensation phenomena in which the ‘change in standard free energy of solution AG" is very small and often close to er0 because of compensation between the enthalpy and entropy terms AHP and TAS" Moreover, in many isothermal processes, thereisa lincar relationship between A/T ‘and AMS (See Box 12.3). This occurs in a family of compounds with @ variable chemical structure, eg. the series of alkylammonium ions when their ionization «energy isconoerned (Fig. 12.5). It also occurs for a given molecule whose solubility is measured in a mixture of water and organic solvent Mm)dl 12.3. HYDRATION OF AMPHIPHILIC MOLECULES am Box 12.3 Study of compensation phenomena Inall eases, the slope of he straight line obtained when A/ is plotted against AS defines a compensation remperature T.. Classically. we have Se AS*=a5+ f (aqnar : and we then se that linear relationship between the two quantities is only possible if AG, = 0. This could occur by chance, but has no general signif canes, We are therefore ld to distinguish between the chemical part of ALP and AS* denoted conventionally by the subscript‘ and the solvation part denoted by the subscript ‘Yat the same time ealizng that an experimental determination ofthe two parts not easy. A linear relationship is assumed ‘soem afar to he sat often = AH = 0+ TAS; | From the definition of Gand replacing AIF by AHfg-+AzHG and AS" by ASE-+AS%, we obtain: ] a AG sa FAM-TAS;-(T-T)AS, | Thequamtitya! = a+ AHj-T.ASzcanbeintroducedsince AMS and AS? are assumed tobe almost constant in given series, and hence ees AG’ =a!-(T-T,)AS,-(T-T.JAS, a + TASS; + AS) - TAS; + AS) ‘Underlying this ad hoe explanation of compensation phenomena, there must be tn interpretation using an equilibrium between lwo states of water Wy and W2 (e.g. to different types of order for the water molecules around a solvated mol- cecule). We should then have tr (AH/AS) for the transition IW > Ws ‘The observed compensation temperature would essentially measure a type of transition between two structures of bound water. 123.3. Micellar organization ‘Assoon as the hydrophobic cain is ong enough, new propetis appear. Beyond a certain concentration of amphiphiles in water « new typeof orders established inthesolutonin the form of micelles, consisting offuey well-defined aggregates of «\cernin number of amphiphilic molecules. The limiting concentration is known ‘the ertea miele concentravion (CMC) (Fi. 12.6). Two opposing forces come 111)mm MOLECULAR BIOPHYSICS Concentration ‘of monomer form ome (@) Total concentration of amphiphilic molecules, Monomer Micelle Concentration of ‘monomer or micelle Figure 12.8 The emaxgenoeoftne ‘gar formnthnereasing concerto: (2) dntion ofthe CMO: {8} tue variaten of moromer and (b) Total concentration of amphiphilic molecules rosie concentrators {nto playin building the micelle (Tanford, 1973) (see Box 12.4): (1) am airracrive van der Waals force between aliphatic chains tending to draw them together within a volume to prevent them dissolving individually; (2) repre force between polar heads, which aréQutside the volume and create a hydrophilic interface with the water. The shape andsize of miele willdepend onthe area covered bythe polarheads, il. om the ratio between the total surface area ofthe micelle and the number of polar heads. There are in general three types of micellar goometry: spherical, cylindrical and double-layer (Fig. 12.7). The morphology and stability of micellar structure depend onthe numberof carbon atoms in the hydrophobic chan and on the number of chains inside the Iie In broad terms, the chain will be almost entirely within the “ntnnal medium’ ofthe micelle, which ean be compared with «guid hydrocarbon ie. tedium with low dieletrie constant. ‘A more quantitative treatmentasin Box 124 enables us to defineanaren forthe polar head depending on the shape ofthe micel he por head area decreases se pas from thesphercl to the planar double layer. Moreover, Box 124shows that the maximum numberof chains Nx packed ina spherical micelle vries asthe quire ofthe numberof carbon atoms nd for 1 = 12Forexample Np 36. In order to pack in the greatest mimber of chains. themicelehasto become a prolateelisoidwithitsminoraxisstilequalt L. After that there isa sudden change to one ofthe other wo shapes, elindreal or double layer which in princplecan accommodate an nfinitenumber of hain. Thechoice |,ll 12.3 HYDRATION OF AMPHIPHILIC MOLECULES 273 be Fe Spherical ‘Cylindrical Double layer aignizaton, 244 Figure 12.8 Georty ola molt chain Box 12.4 Micellar structure ‘This argument follows Cantor and Schimmel (1980). Let’ be the number ‘of carbon atoms inside the micelle. The maximum length of he chain Ly in Fie. 128isgiven by 3 La(A) = 2.1 +063 + 12660/—1) where, on the right-hand side, he Fist term (2.1) represents hindrance of the semi Ctandteseead a 9) staineC- Caste proneton ‘othe axis This equation simplifies to IniA) = 1S + 1.2601 ao ‘For large enough 1, eqn | can be replaced bya relationship ofthe form Lqcmn. Since the stability of « micelle depends on an equilibrium between _twwo opposing forces, the area allocated to each polar head must be caleulated separately foreach overallshaps, However, there isalso relationship between ‘number N ofchainsin the micelleeach containing’ carbonatomsandthe volume ¥ of the micelle This takes the form. | : = . 4 ! since Nis directly proportional to the mass ofthe micelle "spherical micelle of radius J (assumed 0) Tn this ease: ss zl total surfice area = dr? = trae”? e vohime — F-=4rL/3 = 4ra'/3 From eqns and 4: eral (39274 MOLECULAR BIOPHYSICS between these two depends on therepulsive forces between the charged polarheads, hich inereaseas we go from theeylinder to thedoublelayersinee thearea per polar head is halved. ‘When to hydrophobic chains are bound to the polar head, the double-layer {orm is favoured. This is what happens with natural or artificial membranes con- taining phospholipid chains, ‘The material in Boxes 12.5 and 12.6 will help in understanding the thermo- ‘dynamics of micelles. i)i 12.3 HYDRATION OF AMPHIPHILIC MOLECULES 275 Box 125. Transfer of a hydrocarbon from water tothe micelle The tuctre a aliphatic chain inside the celles ery sina to that of __ liquid hydrocarbon, with only a smallextra constraint near the polarheads.To | see this, we stp consider te tanfer ofa bydrophobiceompound between | water and the miele To a st approximation, the ntenorof the mises | ssumed to bean deal solution ofthe compound, for which : j Heaate~1h, = RTV0(Su/Ynicte) | where yay 8 the chemical potential of the amphiphile molecule in the | micellar agerezate | With alkanes, we obtain a inear relationship Mince = 1934711, 8 oe eee aqueous soltion. "eentropy andentay Changes are ofthe sam oder of magnitude, The | fhidity ofthe micellar intvir can also be determined by physical measure- | ments by introducing either « Muorescent probe to follow the decay of flao- ‘rescence with time or a spin label to follow the linewidth of electron spin le formation | Box 12.6 Thermodynamics of mi | The treatment in this box follows Tanford (1973). We wish to calculate the | chemical potential of an amphiphilic molecule in the micellar phase, cen | with respect to the aqueous phase, yg. Ifwe write Vn“ = REIN A + RT Inf ‘we neglect the contribution of the micelle~solvent mixing entropy tothe free ‘nergy. A micellecan be considered as dstinet thermodysamicentity because itis found experimentally to be formed and to be stable only with a rainimum ‘number of amphiphilic molecules. If te size ofthe micelle is ignored and fit ‘contains mamphiphific molecules on average. theeratiecontribution per mole ‘of the amphiphile i Im times the contsibution per mole of micelle. Since the ‘molar fraction Nyce 0 amphiphiles incorporatedin the micelle ism times the ‘molar fraction of micelles, we have that Hits = Hite + (RT) (Xi) Sing, at equim: ome = fr = 16+ RTI Xy + RT ve obtain asf = RT In Xa + RT Inf {RT/) Ise) ‘Theconcept ofa CMC i. acritical value of theconcentration above which “only micelles exist, i only valid in the fit when mse. Ia fact, for large 1 (0276 MOLECULAR BIOPHYSICS | enough my theres almost a phase transition between the molecular solution and the micall since it occurs oversuch a narrow range of concentrations (See Fig. 12.6). Under these conditions. Yq could be replaced by [CMC athe i ele bing re) This esas a _ Forsolutonsof surfactants dhe CMCisa physi quantity asimportantas ‘the melting point or boiling point is for a pute liguid. 1 ean be z "experimentally in many ways: dye binding letrochemical methods, optical ‘orfluorescence,