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S H White

Publications and source records attributed to S H White.

At least 19 recordsLinked to original sources
PubMed1992

Partitioning of tryptophan side-chain analogs between water and cyclohexane.

We have measured the partitioning of the tryptophan side-chain analogs 3-methylindole and N-methylindole between water and cyclohexane over the temperature range 8-55 degrees C to investigate the relative contribution of the imine-NH- to the free energy of transfer. We take advantage of the fact that the indole imine nitrogen is blocked by a methyl group in N-methylindole. Unlike previous studies, we take into account the water present in the cyclohexane phase. Free energies of partitioning were calculated using mole-fraction, volume-fraction, and Flory-Huggins-corrected volume-fraction partition coefficients [De Young, L. R., & Dill, K. A. (1990) J. Phys. Chem. 94, 801-809; Sharp, K. A., Nicholls, A., Friedman, R., & Honig, B. (1991) Biochemistry 30, 9686-9697]. These approaches account for configurational entropy changes in different ways and thus lead to different values for the calculated free energies of transfer. There is a 2-3-fold difference in the free energies calculated from our measurements, using the different units. Independent of units, the partitioning of both compounds involves identical entropy changes. However, 3-methylindole has an additional unfavorable enthalpic contribution to partitioning into cyclohexane of +1.6 kcal/mol (independent of units) which is presumably the cost of removing the indole -NH- group from water and transferring it to cyclohexane. In cyclohexane, 3-methylindole forms hydrogen bonds with water that cause water to copartition into cyclohexane with the solute. A method is described which allows the partitioning process to be examined independent of subsequent interactions with water in the solvent.

Chemical Phenomena↗
PubMed1992

Amino acid preferences of small proteins. Implications for protein stability and evolution.

The dependence of amino acid frequency on sequence length has been examined for the 20 natural amino acids using a set of 2275 protein sequences with little sequence identity. As expected, the frequency of cysteine increases dramatically for sequences shorter than 100 amino acids with a length-dependence that corresponds to an average of two Cys per sequence independent of length. Surprisingly dramatic changes were also observed for the frequencies of arginine, lysine, aspartic acid, and glutamic acid: Arg and Lys frequencies increase for short sequences whereas Asp and Glu frequencies decrease. These changes do not appear to be due to an over-abundance of DNA- and membrane-binding proteins in the database and may, therefore, be related to protein stability. Possible stabilizing mechanisms include increased hydrogen bonding by Arg and increased hydrophobic stabilization due to the amphiphilic character of Arg and Lys. These observations suggest that amino acid composition played an important role in the evolution of small proteins.

Amino Acid Sequence↗
PubMed1992

Structure of a fluid dioleoylphosphatidylcholine bilayer determined by joint refinement of x-ray and neutron diffraction data. II. Distribution and packing of terminal methyl groups.

We continue in this paper the presentation of theoretical and experimental methods for the joint refinement of neutron and x-ray lamellar diffraction data for the analysis of fluid (L alpha phase) bilayer structure (Wiener, M. C., and S. H. White. 1991 a, b, c. Biophys. J. 59:162-173 and 174-185; Biochemistry. 30:6997-7008; Wiener, M. C., G. I. King, and S. H. White. Biophys. J. 60: 568-576). We show how to obtain the distribution and packing of the terminal methyls in the interior of a fluid dioleoylphosphatidylcholine bilayer (66% RH) by combining x-ray and neutron scattering-length transbilayer profiles with no a priori assumptions about the functional form of the distribution. We find that the methyls can be represented by a Gaussian function with 1/e-halfwidth of 2.95 +/- 0.28 A situated at the bilayer center. There is substantial mixing of the methyls and methylenes in the bilayer center. The Gaussian representation of the methyl distribution is narrower and has a different shape than predicted by several simulations of fluid bilayers (Gruen, D. W. R., and E. H. B. de Lacey. 1984. Surfactants in Solution, Vol. 1. Plenum Publishing Corp., New York. 279-306; de Loof, H., et al. 1991. Biochemistry. 30:2099-2133) but this may be due to the smaller area/lipid of our experiments and the presence of the double-bonds. Determination of the absolute specific volume of DOPC and an analysis of bulk alkane volumetric data over a range of hydrostatic pressures lead to estimates of methylene and methyl volumes at the bilayer center of 27 +/- 1 A3 and 57.2 +/- 3.6 A3, respectively. This result provides direct confirmation of the common assumption that the molecular packing of methyl and methylene groups in bilayers is the same as in bulk liquid alkanes.

Biophysical Phenomena↗
PubMed1992

Structure of a fluid dioleoylphosphatidylcholine bilayer determined by joint refinement of x-ray and neutron diffraction data. III. Complete structure.

We present in this paper the complete structure of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) in the L alpha phase (66% RH, 23 degrees C) obtained by the joint refinement of neutron and x-ray lamellar diffraction data. The structural details obtained have previously required a large number of neutron diffraction experiments, using numerous specifically-deuterated phospholipid isomorphs (Büldt et al., 1978. Nature (Lond.). 271:182-184). The joint-refinement approach minimizes specific deuteration by utilizing independent neutron and x-ray data sets. The method yields a quasimolecular structure consisting of a series of multiatomic fragments that are each represented by one or several Gaussian distributions whose positions and widths can be determined to within 0.06 to 0.52 A exclusive of the methylene region. The image of DOPC at 66% RH (5.36 +/- 0.08 waters per lipid) is consistent with many aspects of bilayer structure previously determined by structural and spectroscopic studies. The most striking feature of the structure is the large amount of transbilayer thermal motion suggested by the widths and overlaps of the Gaussian envelopes of the quasimolecular fragments. We discuss the "dynamic bilayer thickness" which describes the minimum effective thickness of the hydrocarbon permeability barrier in terms of the thermal motion of the water. A gradient of thermal motion exists that increases in either direction away from the glycerol backbone which is the most constrained portion of the bilayer. The steric interactions between headgroups of apposed bilayers, expected at the hydration level of our experiments, are clearly revealed. A useful consequence of the quasimolecular structure is that average boundaries within bilayers calculated using composition and volumetric data and ad hoc assumptions can be related to the positions of the principal structural groups. Several measures of "bilayer thickness" in common use can be identified as the positions of the cholines for Luzzati's d1 (Luzzati and Husson. 1962. J. Cell Biol. 12:207-219) and the glycerols for Small's dL (Small. 1967. J. Lipid Res. 8:551-556). We do not know if these relations will be true at other hydrations or for other lipids. Of particular interest is the fact that the position of the carbonyl groups marks the average hydrocarbon/headgroup boundary. It must be emphasized, however, that this region of the bilayer must be generally characterized as one of tumultuous chemical heterogeneity because of the thermal motion of the bilayer.

Biophysical Phenomena↗
PubMed1991

Transbilayer distribution of bromine in fluid bilayers containing a specifically brominated analogue of dioleoylphosphatidylcholine.

We describe in this paper the transbilayer distribution of the bromines of the specifically halogenated phospholipid 1-oleoyl-2-(9,10-dibromostearoyl)-sn-glycero-3- phosphocholine (OBPC). The distribution was determined by X-ray diffraction of oriented multilayers of mixtures of OBPC and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) at 66% relative humidity by the general approach of Franks et al. (1978) [Nature 276, 530-532]. The bromine distribution of OBPC in the fluid L alpha phase is described accurately by a pair of Gaussian functions located 7.97 +/- 0.27 A from the center of the bilayer with l/e half-widths of 4.96 +/- 0.62 A. We find that OBPC bilayers are accurately described as DOPC bilayers with an additional bromine distribution centered at the position of the double bond of DOPC and conclude that OBPC is an excellent structural isomorph for DOPC under the conditions of these experiments. The distribution obtained is the complete and fully resolved transbilayer image of the halogen label because the broad distribution of the bromines is due entirely to thermal disorder and not to experimental limitations [Wiener, M. C., & White, S. H. (1991a) Biophys. J. 59, 162-173]. The observed width of the bromine distribution indicates that virtually all of the hydrocarbon interior is accessible to the bromines. The distance between the bromine/double-bond position and the headgroup phosphate position was determined from one-dimensional Patterson maps and found to be approximately 12 A. The application of accurately determined bromine distributions to the quantitative interpretation of fluorescence quenching experiments is discussed. A method for the self-consistent global analysis of diffraction data from mixtures that permits the use of data sets with different instrumental scale factors is developed in an Appendix.

Bromine↗
PubMed1991

Properties of Asp212----Asn bacteriorhodopsin suggest that Asp212 and Asp85 both participate in a counterion and proton acceptor complex near the Schiff base.

The gene coding for bacteriorhodopsin was modified in vitro to replace Asp212 with asparagine and expressed in Halobacterium halobium. X-ray diffraction measurements showed that the major lattice dimension of purple membrane containing the mutated bacteriorhodopsin was the same as wild type. At pH greater than 7, the Asp212----Asn chromophore was blue (absorption maximum at 585 nm) and exhibited a photocycle containing only the intermediates K and L, i.e. a reaction sequence very similar to that of wild-type bacteriorhodopsin at pH less than 3 and the blue form of the Asp85----Glu protein at pH less than 9. Since in the latter cases these effects are attributed to protonation of residue 85, it now appears that removal of the carboxylate of Asp212 has similar consequences as removing the carboxylate of Asp85. However, an important difference is that only Asp85 affects the pKa of the Schiff base. At pH less than 7, the Asp212----Asn protein was purple (absorption maximum at 569 nm) but photoexcitation produced only 15% of the normal amount of M and the transport activity was partial. The reactions of the blue and purple forms after photoexcitation are both quantitatively accounted for by a proposed scheme, K in equilibrium with L1 in equilibrium with L2----BR, but with the addition of an L1 in equilibrium with M reaction with unfavorable pKa for Schiff base deprotonation in the purple form. The latter hinders the transient accumulation of M, and the consequent branching at L1 allows only partial proton transport activity. The results are consistent with the existence of a complex counterion for the Schiff base proposed earlier (De Groot, H. J. M., Harbison, G. S., Herzfeld, J., and Griffin, R. G. (1989) Biochemistry 28, 3346-3353) and suggest that Asp85, Asp212, and at least one other protonable residue participate in it.

Asparagine↗
PubMed1991

Structure of a fluid dioleoylphosphatidylcholine bilayer determined by joint refinement of x-ray and neutron diffraction data. I. Scaling of neutron data and the distributions of double bonds and water.

We described in two previous papers a method for the joint refinement of the structure of fluid bilayers using neutron and x-ray diffraction data (Wiener, M. C., and S. H. White 1991a, b. Biophys. J. 59: 162-173 and 174-185). An essential part of the method is the appropriate scaling of the diffraction data. Here we describe the scaling of the neutron data and the determination of the transbilayer distribution of double bonds in liquid-crystalline (L alpha phase) phospholipid bilayers of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC). The distribution was determined by neutron diffraction of oriented multilayers (66% RH) of DOPC specifically deuterated at the 9- and 10-position of both acyl chains. The double-bond distribution is described accurately by a pair of Gaussian functions each located at a position Zcc = 7.88 +/- 0.09 A from the bilayer center with 1/e-halfwidths of Acc = 4.29 +/- 0.16 A. Previously, we determined the transbilayer distribution of bromine atoms in a specifically halogenated lipid, 1-oleoyl-2-9,10-dibromostearoyl-sn-glycero-3-phosphocholine (OBPC), and showed it to be an isomorphous replacement for DOPC (Wiener, M. C., and S. H. White, 1991c. Biochemistry. In press). A comparison of the double-bond and bromine profiles indicates that the positions of the centers of the deuterated double bond and the brominated methylene Gaussian distributions are equal within experimental error and that each label undergoes similar average thermal motions with respect to the bilayer normal. The observation that the average position of a label on both acyl chains (the deuterated double bonds) is similar to the average position of a label on the 2-chain alone (the brominated methylenes) indicates that the maximum separation along the bilayer normal between the double bonds of the acyl chains is 1 A or less. The fully-resolved transbilayer water distribution, previously determined at lower resolution (Jacobs, R. E., and S. H. White. 1989. Biochemistry. 28:3421-3437), was obtained from the analysis of neutron diffraction data of DOPC hydrated with a D20/H20 mixture. The water distribution is described accurately by a pair of Gaussian functions each located at a position Zw = 22.51 +/- 0.77 A from the bilayer center with 1/e-half widths of Aw = 4.63 +/- 0.48A. We present the relative absolute neutron and x-ray structure factors of DOPC at 66% RH that will be used to solve the complete structure of DOPC which will be presented in a later paper of this series.

Lipid Bilayers↗
PubMed1991

Fluid bilayer structure determination by the combined use of x-ray and neutron diffraction. I. Fluid bilayer models and the limits of resolution.

This is the first in a series of papers concerned with methods for the determination of the structures of fluid phospholipid bilayers in the liquid-crystalline (L alpha) phase. The basic approach is the joint refinement of quasimolecular models (King and White, 1986. Biophys. J. 49:1047-1054) using x-ray and neutron diffraction data. We present here (a) the rationale for quasimolecular models, (b) the nature of the resolution problem for thermally disordered bilayers, and (c) an analysis of the resolution of experiments in which Gaussian functions are used to describe the distribution of submolecular components. We show that multilamellar liquid-crystalline bilayers are best described by the convolution of a perfect lattice function with a thermally disordered bilayer unit cell. Lamellar diffraction measurements on such a system generally yield only 5-10 orders of diffraction data from which transbilayer profiles of the unit cell can be constructed. The canonical resolution of these transbilayer profiles, defined as the Bragg spacing divided by the index of the highest recorded diffraction order, is typically 5-10 A. Using simple model calculations, we show that the canonical resolution is a measure of the widths of the distributions of constituents of the unit cell rather than a measure of the spatial separation of the distributions. The widths provide a measure of the thermal motion of the bilayer constituents which can be described by Gaussian functions. The equilibrium positions of the centers of the distributions can be determined with a precision of 0.1-0.5 A based upon typical experimental errors.

Lipid Bilayers↗
PubMed1991

Fluid bilayer structure determination by the combined use of x-ray and neutron diffraction. II. "Composition-space" refinement method.

This is the second of two papers describing a method for the joint refinement of the structure of fluid bilayers using x-ray and neutron diffraction data. We showed in the first paper (Wiener, M. C., and S. H. White. 1990. Biophys. J. 59:162-173) that fluid bilayers generally consist of a nearly perfect lattice of thermally disordered unit cells and that the canonical resolution d/hmax is a measure of the widths of quasimolecular components represented by simple Gaussian functions. The thermal disorder makes possible a "composition space" representation in which the quasimolecular Gaussian distributions describe the number or probability of occupancy per unit length across the width of the bilayer of each component. This representation permits the joint refinement of neutron and x-ray lamellar diffraction data by means of a single quasimolecular structure that is fit simultaneously to both diffraction data sets. Scaling of each component by the appropriate neutron or x-ray scattering length maps the composition space profile to the appropriate scattering length space for comparison to experimental data. Other extensive properties, such as mass, can also be obtained by an appropriate scaling of the refined composition space structure. Based upon simple bilayer models involving crystal and liquid crystal structural information, we estimate that a fluid bilayer with hmax observed diffraction orders will be accurately represented by a structure with approximately hmax quasimolecular components. Strategies for assignment of quasimolecular components are demonstrated through detailed parsing of a phospholipid molecule based upon the one-dimensional projection of the crystal structure of dimyristoylphosphatidylcholine. Finally, we discuss in detail the number of experimental variables required for the composition space joint refinement. We find fluid bilayer structures to be marginally determined by the experimental data. The analysis of errors, which takes on particular importance under these circumstances, is also discussed.

Lipid Bilayers↗
PubMed1991

Membrane structures in normal and essential fatty acid-deficient stratum corneum: characterization by ruthenium tetroxide staining and x-ray diffraction.

Despite the importance of intercellular lamellar bilayers for stratum corneum (SC) barrier function, knowledge about the structure of these bilayers is limited due to their poor visualization and/or retention. Whereas substitution of ruthenium tetroxide (RuO4) for osmium tetroxide fixation provides clear images of these bilayers, the usefulness of RuO4 has been limited by its slow penetration and cytotoxicity. Utilizing a new fixation protocol for RuO4, we obtained clear images of lamellar domains at all levels of murine SC. Computer-aided image reconstructions demonstrated a lamellar spacing of 129 +/- 2 A, which agreed with x-ray diffraction data from parallel, unfixed samples (131 +/- 2 A), a spacing not affected by hydration. Furthermore, novel structures were seen in the intercellular spaces of normal SC. Finally, in murine essential fatty acid deficiency (EFAD), the overall lamellar spacing is comparable to normal [127 +/- 7 A by computer transform vs. 131.9 +/- 2 A (hydrated) and 129.6 +/- 2.2 A (dry) by x-ray diffraction]. Yet, these domains are structurally abnormal, displaying regions with either an excess or absence of lamellae. The new RuO4 protocol provides quantitative information about SC lamellar dimensions and morphologic abnormalities in bilayer distribution and substructure in EFAD stratum corneum that are not detected by either x-ray diffraction or computer-aided image reconstruction. Thus, the barrier abnormality in EFAD stratum corneum can be ascribed either to focal depletion of lamellae or abnormalities in lamellar substructure.

Animals↗
PubMed1991

Sagittal plane laxity following knee arthroplasty.

We measured the sagittal laxity in 70 knee replacements at least six months after surgery, using a KT 1000 arthrometer. With an unconstrained prosthesis (the Oxford meniscal knee) anteroposterior stability was normal in joints known to have intact cruciate ligaments. There was increased laxity in those which lacked an anterior cruciate ligament. In knees with an intact anterior cruciate ligament, sagittal laxity did not increase with time.

Anterior Cruciate Ligament↗
PubMed1991

Anteromedial osteoarthritis of the knee.

Medial tibial plateaux excised during 46 unicompartmental arthroplasties for osteoarthritis were collected and photographed. The anterior cruciate ligament was intact in all joints. In every case the cartilage and bone erosion was centred anteriorly on the plateau and the posterior cartilage was intact. The site of the lesion and the intact state of the cruciate ligaments taken together explain why varus deformity was observed only in the extended knee, and why the deformity was correctable and had not become fixed. Failure of the anterior cruciate ligament may allow the erosion to extend posteriorly, producing fixed varus deformity and leading to degeneration of the lateral compartment. Anteromedial osteoarthritis is a distinct clinicopathological entity; its radiographic features enable it to be diagnosed from lateral radiographs; its anatomical features render it suitable for treatment by unicompartmental arthroplasty.

Age Factors↗
PubMed1991

Axial movement and tibial fractures. A controlled randomised trial of treatment.

Diaphyseal fractures of the tibia in 80 patients were treated by external skeletal fixation using a unilateral frame, either in a fixed mode or in a mode which allowed the application of a small amount of predominantly axial micromovement. Patients were allocated to each regime by random selection. Fracture healing was assessed clinically, radiologically and by measurement of the mechanical stiffness of the fracture. Both clinical and mechanical healing were enhanced in the group subjected to micromovement, compared to those treated with frames in a fixed mode possessing an overall stiffness similar to that of others in common clinical use. The differences in healing time were statistically significant and independently related to the treatment method. There was no difference in complication rates between treatment groups.

Adolescent↗
PubMed1991

The importance of delay in distraction of osteotomies.

The factor of timing of the commencement of distraction of an osteotomy is evaluated. Experimental osteotomies subjected to immediate distraction result in the production of a small volume of callus with deficient vascularity. Delay alters this response, and there is a more cellular osteogenic reaction with proliferative vascularization during lengthening. A delay is advocated in all gradual leg lengthening procedures, in progressive correction of deformity controlled by external skeletal fixation following osteotomy, and in bone transport procedures. The period of delay should be carefully determined according to each individual case.

Animals↗
PubMed1991

Closed suction drainage following knee arthroplasty. Effectiveness and risks.

A prospective investigation was performed to determine when to remove a suction drain following total knee arthroplasty (TKA). Forty-one TKAs were randomly allocated to closed suction drainage for either 24 or 48 hours. The drain was removed and the tip was cut off and processed by a method giving quantitative cultures. In the 48-hour group, 85% of the total volume was drained during the first 24 hours. During the following 24-hour period, a mean volume of only 50 ml was drained. No organism was isolated from cultures of drain tips sampled at 24 hours. However, at 48 hours, 25% of the drain tips yielded light growths of coagulase-negative staphylococci (four drain tips) and Staphylococcus aureus (one drain tip). Clinical evaluations of wound healing were comparable in the two groups. Clearly, nothing is to be gained by continuing drainage beyond 24 hours. If drainage is maintained for longer periods, there is an increased risk of contamination by bacteria.

Equipment Contamination↗
PubMed1990

Observations concerning topology and locations of helix ends of membrane proteins of known structure.

Hydropathy plots of amino acid sequences reveal the approximate locations of the transbilayer helices of membrane proteins of known structure and are thus used to predict the helices of proteins of unknown structure. Because the three-dimensional structures of membrane proteins are difficult to obtain, it is important to be able to extract as much information as possible from hydropathy plots. We describe an "augmented" hydropathy plot analysis of the three membrane proteins of known structure, which should be useful for the systematic examination and comparison of membrane proteins of unknown structure. The sliding-window analysis utilizes the floating interfacial hydrophobicity scale [IFH(h)] of Jacobs and White (Jacobs, R.E., White, S. H., 1989. Biochemistry 28:3421-3437) and the reverse-turn (RT) frequencies of Levitt (Levitt, M., 1977, Biochemistry 17:4277-4285). The IFH(h) scale allows one to examine the consequences of different assumptions about the average hydrogen bond status (h = 0 to 1) of polar side chains. Hydrophobicity plots of the three proteins show that (i) the intracellular helix-connecting links and chain ends can be distinguished from the extracellular ones and (ii) the main peaks of hydrophobicity are bounded by minor ones which bracket the helix ends. RT frequency plots show that (iii) the centers of helices are usually very close to wide-window minima of average RT frequency and (iv) helices are always bounded by narrow-window maxima of average RT frequency. The analysis suggests that side-chain hydrogen bonding with membrane components during folding may play a key role in insertion.

Amino Acid Sequence↗