Massive in Silico Study of Noble Gas Binding to the Structural Proteome.

Noble gases are chemically inert, and it was therefore thought they would have little effect on biology. Paradoxically, it was found that they do exhibit a wide range of biological effects, many of which are target-specific and potentially useful and some of which have been demonstrated in vivo. The underlying mechanisms by which useful pharmacology, such as tissue and neuroprotection, anti-addiction effects, and analgesia, is elicited are relatively unexplored. Experiments to probe the interactions of noble gases with specific proteins are more difficult with gases than those with other chemicals. It is clearly impractical to conduct the large number of gas-protein experiments required to gain a complete picture of noble gas biology. Given the simplicity of atoms as ligands, in silico methods provide an opportunity to gain insight into which noble gas-protein interactions are worthy of further experimental or advanced computational investigation. Our previous validation studies showed that in silico methods can accurately predict experimentally determined noble gas binding sites in X-ray structures of proteins. Here, we summarize the largest reported in silico reverse docking study involving 127 854 protein structures and the five nonradioactive noble gases. We describe how these computational screening methods are implemented, summarize the main types of interactions that occur between noble gases and target proteins, describe how the massive data set that this study generated can be analyzed (freely available at group18.csiro.au), and provide the NDMA receptor as an example of how these data can be used to understand the molecular pharmacology underlying the biology of the noble gases. We encourage chemical biologists to access the data and use them to expand the knowledge base of noble gas pharmacology, and to use this information, together with more efficient delivery systems, to develop "atomic drugs" that can fully exploit their considerable and relatively unexplored potential in medicine.

Pharmacokinetics of tobramycin following intravenous, intramuscular, and intra-articular administration in healthy horses.

The objectives of this study were to examine the pharmacokinetics of tobramycin in the horse following intravenous (IV), intramuscular (IM), and intra-articular (IA) administration. Six mares received 4 mg/kg tobramycin IV, IM, and IV with concurrent IA administration (IV+IA) in a randomized 3-way crossover design. A washout period of at least 7 days was allotted between experiments. After IV administration, the volume of distribution, clearance, and half-life were 0.18 ± 0.04 L/kg, 1.18 ± 0.32 mL·kg/min, and 4.61 ± 1.10 h, respectively. Concurrent IA administration could not be demonstrated to influence IV pharmacokinetics. The mean maximum plasma concentration (Cmax ) after IM administration was 18.24 ± 9.23 µg/mL at 1.0 h (range 1.0-2.0 h), with a mean bioavailability of 81.22 ± 44.05%. Intramuscular administration was well tolerated, despite the high volume of drug administered (50 mL per 500 kg horse). Trough concentrations at 24 h were below 2 µg/mL in all horses after all routes of administration. Specifically, trough concentrations at 24 h were 0.04 ± 0.01 µg/mL for the IV route, 0.04 ± 0.02 µg/mL for the IV/IA route, and 0.02 ± 0.02 for the IM route. An additional six mares received IA administration of 240 mg tobramycin. Synovial fluid concentrations were 3056.47 ± 1310.89 µg/mL at 30 min after administration, and they persisted for up to 48 h with concentrations of 14.80 ± 7.47 µg/mL. Tobramycin IA resulted in a mild chemical synovitis as evidenced by an increase in synovial fluid cell count and total protein, but appeared to be safe for administration. Monte Carlo simulations suggest that tobramycin would be effective against bacteria with a minimum inhibitory concentration (MIC) of 2 µg/mL for IV administration and 1 µg/mL for IM administration based on Cmax :MIC of 10.

Endoscopic anatomy of the cervical vertebral canal in the horse: a cadaver study.

REASON FOR PERFORMING STUDY: Localisation of spinal cord compression in horses with cervical vertebral stenotic myelopathy is inexact. Vertebral canal endoscopy has been used in man to localise spinal cord lesions and has the potential to become a useful diagnostic technique in horses. OBJECTIVE: To establish a surgical approach via the atlanto-occipital space to the cervical vertebral canal in equine cadavers and describe the endoscopic anatomy of the cervical epidural and subarachnoid spaces. METHODS: The cadavers of 25 mature horses were used to assess 3 surgical methods to approach the cervical vertebral canal, including 2 minimally invasive and one open technique. Once the approach had been made, a flexible videoendoscope was inserted into the epidural space (epiduroscopy) or the subarachnoid space (myeloscopy) and advanced caudally until the intervertebral space between C7 and T1 was reached. RESULTS: The epidural and subarachnoid spaces could not be accessed reliably using the minimally invasive techniques. Furthermore, damage to the nervous tissues was a frequent complication with these procedures. The open approach allowed successful insertion of the videoendoscope into the epidural and subarachnoid spaces in all horses and no inadvertent damage was observed. Anatomical structures that could be seen in the epidural space included the dura mater, nerve roots, fat and the ventral internal vertebral venous plexus. In the subarachnoid space, the spinal cord, nerve roots, blood vessels, denticulate ligaments and external branch of the accessory nerve were seen. CONCLUSIONS: Using the open approach, epiduroscopy and myeloscopy over the entire length of the cervical vertebral canal are possible in the mature horse. POTENTIAL RELEVANCE: Cervical vertebral canal endoscopy may become a valuable tool to localise the site of spinal cord injury in horses with cervical vertebral stenotic myelopathy and could aid in the diagnosis of other diseases of the cervical spinal cord.

Comparison of conventional and alternative arthroscopic approaches to the palmar/plantar pouch of the equine distal interphalangeal joint.

REASONS FOR PERFORMING THE STUDY: The conventional arthroscopic approach to the palmar/plantar aspect of the distal interphalangeal joint (DIPJ) may result in the inadvertent penetration of the digital flexor tendon sheath (DFTS) and the navicular bursa (NB). This iatrogenic communication would be undesirable subsequent to arthroscopic lavage of a septic DIPJ. HYPOTHESIS: A lateral/medial approach to the palmar/plantar aspect of the DIPJ will result in a significantly lower rate of inadvertent penetration of the DFTS and NB, whilst still providing adequate intra-articular evaluation. METHODS: The conventional palmar/plantar approach or a novel lateral/medial approach to the DIPJ was performed on cadaver fore- and hindlimbs (30 limbs/approach). Subsequently, India ink was injected into the dorsal pouch of the DIPJ, and the DFTS (n = 60) and NB (n = 20) were examined for the presence/absence of ink. In addition, observations of the number of attempts made to access the joint, evidence of iatrogenic intra-articular trauma and occurrence of incomplete visualisation of the palmar/plantar pouch were recorded. RESULTS: With the conventional approach, DFTS penetration was noted in 18/30 (60%) of the limbs, compared to 1/30 (3.3%) with the lateral/medial approach (P≤0.001). NB penetration was seen in 5/10 limbs with the palmar/plantar approach compared to 0/10 with the lateral/medial approach (P = 0.01). No significant differences were found between the approaches in the number of attempts made to access the joint, the incidence of iatrogenic intra-articular trauma, or the occurrence of incomplete visibility of the palmar/plantar pouch. CONCLUSIONS: The novel lateral/medial approach to the DIPJ significantly decreases the risk of inadvertent penetration of the DFTS and NB. POTENTIAL RELEVANCE: The novel lateral/medial approach to the DIPJ is an effective technique to gain access to the palmar/plantar pouches, and is particularly advantageous for arthroscopic lavage of a septic DIPJ.

Effects of danofloxacin dosing regimen on gastrointestinal pharmacokinetics and fecal microbiome in steers.

Fluoroquinolones are a class of antimicrobial commonly used in human medicine, and deemed critical by the World Health Organization. Nonetheless, two formulations are approved for the treatment of respiratory disease in beef cattle. The objective of this study was to determine the gastrointestinal pharmacokinetics and impact on enteric bacteria of cattle when receiving one of the two dosing regimens (high: 40 mg/kg SC once or low: 20 mg/kg IM q48hr) of danofloxacin, a commonly utilized synthetic fluoroquinolone in veterinary medicine. Danofloxacin was administered to 12 steers (age 7 months) fitted with intestinal ultrafiltration devices at two different dosing regimens to assess the gastrointestinal pharmacokinetics, the shifts in the gastrointestinal microbiome and the development of resistant bacterial isolates. Our results demonstrated high intestinal penetration of danofloxacin for both dosing groups, as well as, significant differences in MIC values for E. coli and Enterococcus between dosing groups at selected time points over a 38 day period. Danofloxacin treatment consistently resulted in the Euryarchaeota phyla decreasing over time, specifically due to a decrease in Methanobrevibacter. Although microbiome differences were minor between dosing groups, the low dose group had a higher number of isolates with MIC values high enough to cause clinically relevant resistance. This information would help guide veterinarians as to appropriate dosing schemes to minimize the spread of antimicrobial resistance.

The crystal structure of oxylipin-conjugated barley LTP1 highlights the unique plasticity of the hydrophobic cavity of these plant lipid-binding proteins.

The barley lipid transfer protein (LTP1) adducted by an alpha-ketol, (9-hydroxy-10-oxo-12(Z)-octadecenoic acid) exhibits an unexpected high lipid transfer activity. The crystal structure of this oxylipin-adducted LTP1, (LTP1b) was determined at 1.8A resolution. The covalently bound oxylipin was partly exposed at the surface of the protein and partly buried within the hydrophobic cavity. The structure of the oxylipidated LTP1 emphasizes the unique plasticity of the hydrophobic cavity of these plant lipid-binding proteins when compared to the other members of the family. The plasticity of the hydrophobic cavity and increase of its surface hydrophobicity induced by the oxylipin account for the improvement of the lipid transfer activity of LTP1b. These observations open new perspectives to explore the different biological functions of LTPs, including their allergenic properties.

Feasibility and safety of lumbosacral epiduroscopy in the standing horse.

REASONS FOR PERFORMING STUDY: The large size of the adult horse prevents the use of advanced imaging modalities in most areas of the axial skeleton, including the lumbosacral vertebral column. Traditional imaging techniques are frequently unable to pinpoint the underlying pathology in horses with caudal back pain. In man, lumbosacral epiduroscopy is used to diagnose and treat subjects with chronic back and leg pain. This technique may close the diagnostic gap in horses with similar clinical signs. OBJECTIVES: To evaluate the safety and feasibility of lumbosacral epiduroscopy in the standing adult horse. STUDY DESIGN: Descriptive, experimental study. METHODS: Seven adult horses weighing 504-578 kg were sedated and restrained in stocks in preparation for aseptic surgery. Vascular dilators of increasing size were inserted cranial to the first moveable vertebra caudal to the sacrum to facilitate a minimally invasive approach into the epidural space. A flexible video-endoscope was introduced and advanced as far as its 60-cm working length permitted. Pre-, intra- and post-operative plasma cortisol samples were collected, and neurological and lameness examinations were performed prior to and during the 2 weeks following the procedure. Post-mortem examinations were conducted in 5 of the 7 horses. RESULTS: Standing lumbosacral epiduroscopy was well tolerated by all horses. The anatomic structures in the epidural space (dura mater, spinal nerve roots, fat and blood vessels) were followed as far cranial as the thoracolumbar region. No complications related to the procedure were noted in the 2-week monitoring period following epiduroscopy. Small, organised haematomas were identified in the sacral epidural space during necropsy in one horse. No abnormalities were seen in the other 4 animals. CONCLUSIONS: Lumbosacral epiduroscopy can be performed safely in sedated standing horses. The procedure may become a valuable diagnostic tool in horses with caudal back or hindlimb pain of unknown origin.

The catalytic site of serine proteinases as a specific binding cavity for xenon.

BACKGROUND: Under moderate pressure, xenon can bind to proteins and form weak but specific interactions. Such protein-xenon complexes can be used as isomorphous derivatives for phase determination in X-ray crystallography. RESULTS: Investigation of the serine proteinase class of enzymes shows that the catalytic triad, the common hydrolytic motif of these enzymes, is a specific binding site for one xenon atom and shows high occupancy at pressures below 12 bar. Complexes of xenon with two different serine proteinases, elastase and collagenase, were analyzed and refined to 2.2 A and 2.5 A resolution, respectively. In both cases, a single xenon atom with a low temperature factor is located in the active site at identical positions. Weak interactions exist with several side chains of conserved amino acids at the active site. Xenon binding does not induce any major changes in the protein structure and, as a consequence, crystals of the xenon complexes are highly isomorphous with the native protein structures. Xenon is also found to bind to the active site of subtilisin Carlsberg, a bacterial serine proteinase, that also has a catalytic triad motif. CONCLUSIONS: As the region around the active site shows conserved structural homology in all serine proteinases, it is anticipated that xenon binding will prove to be a general feature of this class of proteins.

A zipper-like duplex in DNA: the crystal structure of d(GCGAAAGCT) at 2.1 A resolution.

BACKGROUND: The replication origin of the single-stranded (ss)DNA bacteriophage G4 has been proposed to fold into a hairpin loop containing the sequence GCGAAAGC. This sequence comprises a purine-rich motif (GAAA), which also occurs in conserved repetitive sequences of centromeric DNA. ssDNA analogues of these sequences often show exceptional stability which is associated with hairpin loops or unusual duplexes, and may be important in DNA replication and centromere function. Nuclear magnetic resonance (NMR) studies indicate that the GCGAAAGC sequence forms a hairpin loop in solution, while centromere-like repeats dimerise into unusual duplexes. The factors stabilising these unusual secondary structure elements in ssDNA, however, are poorly understood. RESULTS: The nonamer d(GCGAAAGCT) was crystallised as a bromocytosine derivative in the presence of cobalt hexammine. The crystal structure, solved by the multiple wavelength anomalous dispersion (MAD) method at the bromine K-edge, reveals an unexpected zipper-like motif in the middle of a standard B-DNA duplex. Four central adenines, flanked by two sheared G.A mismatches, are intercalated and stacked on top of each other without any interstrand Watson-Crick base pairing. The cobalt hexammine cation appears to participate only in crystal cohesion. CONCLUSIONS: The GAAA consensus sequence can dimerise into a stable zipper-like duplex as well as forming a hairpin loop. The arrangement closes the minor groove and exposes the intercalated, unpaired, adenines to the solvent and DNA-binding proteins. Such a motif, which can transform into a hairpin, should be considered as a structural option in modelling DNA and as a potential binding site, where it could have a role in DNA replication, nuclease resistance, ssDNA genome packaging and centromere function.

Crystal structure of a D-aminopeptidase from Ochrobactrum anthropi, a new member of the 'penicillin-recognizing enzyme' family.

BACKGROUND: beta-Lactam compounds are the most widely used antibiotics. They inactivate bacterial DD-transpeptidases, also called penicillin-binding proteins (PBPs), involved in cell-wall biosynthesis. The most common bacterial resistance mechanism against beta-lactam compounds is the synthesis of beta-lactamases that hydrolyse beta-lactam rings. These enzymes are believed to have evolved from cell-wall DD-peptidases. Understanding the biochemical and mechanistic features of the beta-lactam targets is crucial because of the increasing number of resistant bacteria. DAP is a D-aminopeptidase produced by Ochrobactrum anthropi. It is inhibited by various beta-lactam compounds and shares approximately 25% sequence identity with the R61 DD-carboxypeptidase and the class C beta-lactamases. RESULTS: The crystal structure of DAP has been determined to 1.9 A resolution using the multiple isomorphous replacement (MIR) method. The enzyme folds into three domains, A, B and C. Domain A, which contains conserved catalytic residues, has the classical fold of serine beta-lactamases, whereas domains B and C are both antiparallel eight-stranded beta barrels. A loop of domain C protrudes into the substrate-binding site of the enzyme. CONCLUSIONS: Comparison of the biochemical properties and the structure of DAP with PBPs and serine beta-lactamases shows that although the catalytic site of the enzyme is very similar to that of beta-lactamases, its substrate and inhibitor specificity rests on residues of domain C. DAP is a new member of the family of penicillin-recognizing proteins (PRPs) and, at the present time, its enzymatic specificity is clearly unique.

Epiduroscopy of the lumbosacral vertebral canal in the horse: Technique and endoscopic anatomy.

REASONS FOR PERFORMING STUDY: Back pain is a common cause of gait alterations and poor performance in horses, but the available imaging modalities are frequently insufficient to isolate the underlying pathology. In human patients, epidural endoscopy (epiduroscopy) is successfully used to diagnose and treat challenging cases of lower back pain. Endoscopy of the cervical epidural space has previously been reported in anaesthetised horses. OBJECTIVES: To develop a technique for lumbosacral epiduroscopy in standing horses and to describe the endoscopic anatomy of the lumbosacral epidural space. STUDY DESIGN: Pilot study to assess the feasibility of lumbosacral epiduroscopy in 5 horse cadavers. METHODS: The cadavers of 5 horses, weighing 457-694 kg (mean, 570 kg), were suspended in an upright position. Vascular dilators of increasing size were inserted between the first 2 moveable vertebrae caudal to the sacrum to create a minimally invasive approach into the epidural space. A flexible videoendoscope was introduced and advanced as far cranially as the length of the endoscope permitted. The lumbosacral epidural space underwent gross necropsy examination following the procedure. RESULTS: The endoscope was successfully inserted into the epidural space in all horses. Saline injection through the working channel of the endoscope allowed the following anatomical structures to be seen: dura mater, left and right lumbosacral spinal nerves, cauda equina, epidural fat, connective tissue and blood vessels. Using the 60 cm working length of the endoscope, the epidural space could be examined as far cranial as L3-T18, depending on the size of the horse. No gross damage to epidural neurovascular structures was observed on necropsy examination. CONCLUSION: Lumbosacral epiduroscopy is technically feasible in standing horses and may become a valuable diagnostic tool in horses with caudal back or limb pain of unknown origin. Studies in live horses will be necessary to evaluate the safety of the procedure.

Navigational ultrasound imaging: A novel imaging tool for aiding interventional therapies of equine musculoskeletal injuries.

REASONS FOR PERFORMING STUDY: Navigational ultrasound imaging, also known as fusion imaging, is a novel technology that allows real-time ultrasound imaging to be correlated with a previously acquired computed tomography (CT) or magnetic resonance imaging (MRI) study. It has been used in man to aid interventional therapies and has been shown to be valuable for sampling and assessing lesions diagnosed with MRI or CT that are equivocal on ultrasonography. To date, there are no reports of the use of this modality in veterinary medicine. OBJECTIVES: To assess whether navigational ultrasound imaging can be used to assist commonly performed interventional therapies for the treatment of equine musculoskeletal injuries diagnosed with MRI and determine the appropriateness of regional anatomical landmarks as registration sites. STUDY DESIGN: Retrospective, descriptive clinical study. METHODS: Horses with musculoskeletal injuries of the distal limb diagnosed with MRI scheduled for ultrasound-guided interventional therapies were evaluated (n = 17 horses with a total of 29 lesions). Anatomical landmarks used for image registration for the navigational procedure were documented. Accuracy of lesion location and success of the procedure were assessed subjectively and described using a grading scale. RESULTS: All procedures were accurately registered using regional anatomical landmarks and considered successful based on our criteria. Anatomical landmarks were described for each lesion type. The addition of navigational imaging was considered to greatly aid the procedures in 59% of cases and added information to the remainder of the procedures. The technique was considered to improve the precision of these interventional procedures. CONCLUSIONS: Navigational ultrasound imaging is a complementary imaging modality that can be used for the treatment of equine soft tissue musculoskeletal injuries diagnosed with MRI.

Molecular structure of the lipoamide dehydrogenase domain of a surface antigen from Neisseria meningitidis.

The protein p64k from the surface of the Neisseria meningitidis bacteria has been characterized as a two-domain protein. It contains a dihydrolipoamide dehydrogenase domain of 482 residues, involving a FAD prosthetic group as a cofactor, and a smaller lipoic acid binding domain of 86 residues. The two domains are joined by a flexible segment rich in alanine and proline residues. The structure of the dihydrolipoamide dehydrogenase domain was determined by X-ray diffraction. It was solved by a combination of molecular replacement and multiple isomorphous replacement techniques and refined to 2.7 A resolution. In the crystal, the recombinant p64k mimics the functional homo-dimer by using one of the crystallographic 2-fold axes. The reactive disulphide bridge Cys161-Cys166 is in the oxidised state and the FAD is bound in an extended conformation. This main domain contains the major antigenic determinant of the protein, an extended loop of 32 residues at the surface of the protein. A mis-attribution at residue 553 in the sequence has been detected by inspection of electron density maps and the geometry. However, when compared to the other dihydrolipoamide dehydrogenases, there are some significant differences: (1) an unusual number of cis-proline residues and (2) a new motif built around a 2-fold axis by the sulphur atoms of residues Met558, Cys560 and their symmetry related equivalents.

Crystallization and preliminary X-ray investigation of a recombinant outer membrane protein from Neisseria meningitidis.

A protein constituent of the outer membrane from Neisseria meningitidis (hereafter called P64K) has been crystallized using the hanging drop technique. Crystals are tetragonal with unit cell dimensions a = b = 136.84 A and c = 78.44 A, compatible with a single monomer of 64 kDa in the asymmetric unit. When exposed to high intensity synchrotron radiation, these crystals diffract X-rays to at least 2.9 A resolution, indicating that a high resolution structure analysis is feasible.

The BPTI decamer observed in acidic pH crystal forms pre-exists as a stable species in solution.

Bovine pancreatic trypsin inhibitor (BPTI) crystallizes under acidic pH conditions in the presence of thiocyanate, chloride and sulfate ions, yielding three different polymorphs in P2(1), P6(4)22 and P6(3)22 space groups, respectively. In all three crystal forms, the same decamer is found in the packing (ten BPTI molecules organized through two perpendicular 2-fold and 5-fold axes as a well-defined and compact object) in contrast to the monomeric crystal forms observed at basic pH conditions. The crystallization of BPTI under acidic conditions (pH 4.5) was investigated by small angle X-ray scattering with both under- and supersaturated BPTI solutions. Data showed the oligomerization of BPTI molecules under all investigated conditions. Accordingly, various mixtures of discrete oligomers (n=1 to 10) were considered. Calculated scattering curves were obtained using models based on the crystallographic structures, and the experimental patterns were analyzed as a linear combination of the model curves using a non-linear curve fitting procedure. The results, confirmed by gel filtration experiments, unambiguously demonstrate the co-existence of two different BPTI particles in solution: a monomer and a decamer, with no evidence of any other intermediates. Moreover, using both approaches, the fraction of decamers was found to increase with increasing salt concentration, even beyond the solubility curve. We therefore propose that at acidic pH, BPTI crystallizes following a two step process: decamers are first built in under- and supersaturated solutions, upon which crystal growth proceeds by decamer stacking. Indeed, those BPTI crystals should best be described as "BPTI decamer" crystals.

Structure of tumour necrosis factor by X-ray solution scattering and preliminary studies by single crystal X-ray diffraction.

The structure of tumour necrosis factor has been investigated by X-ray small-angle scattering and X-ray diffraction using synchrotron radiation. The overall radius of gyration is 25.5 A. A plausible model accounting for the scattering curves consists of an elongated trimer with an axial ratio of 3 to 4 and a maximal chord with a lower limit of 80 A. Tumour necrosis factor has been crystallized in a trigonal space group. Our results are in favour of a single trimer in the asymmetric unit. The diffraction extends to 3.5 A.

Crystallization of the MS2 translational repressor alone and complexed to bromouridine.

The coat protein from the MS2 bacteriophage plays a dual role by encapsidating viral RNA and also by binding RNA as a translational repressor. In order to study the isolated dimer in a conformation not influenced by capsid interactions, a mutant molecule was crystallized that is defective in capsid assembly but is an active repressor. The unassembled dimer crystallized in the space group P21212 with a = 76.2, b = 55.7, and c = 28.4 A. In these crystals, monomers were related by twofold symmetry. When this dimer was co-crystallized with 5-bromouridine, crystals formed in space group R3 with a = b = 155.9 A, c = 29.9 A, gamma = 120 degrees; the dimer was the asymmetric unit.

Correct intron splicing generates a new type of a putative zinc-binding domain in a transcriptional activator of Aspergillus nidulans.

alcR is the pathway-specific transcriptional activator of the ethanol regulon in the filamentous fungus, Aspergillus nidulans. The deduced amino acid sequence of a cDNA clone, including the 5' part of the alcR-mRNA, shows that a putative Zn-binding domain of the all-cysteine class, exemplified by GAL4 is present. This structure presents some striking features. At variance with other structures of this class, the binding domain is strongly asymmetrical. Model building indicates that the zinc-binding motif of alcR could adopt an helix-turn-helix structure. We propose that the DNA-binding motif of alcR could participate in two types of DNA-binding structures: the zinc-cluster and the helix-turn-helix.

Angiography contrast agents interact with serine proteinases. The molecular structure of the model system elastase/iohexol.

Pancreatic elastase was co-crystallised with iohexol, a tri-iodo benzenic contrast agent used in angiography analyses. The X-ray analysis of the complex reveals the presence of three molecules of iohexol associated with the proteinase with low occupancy factors. Two iohexol molecules are located in and near the active site of the enzyme and provide a model for explaining the inhibition of the hemostatic system, one of the major and inconvenient side effect associated with these chemicals.

Fatty acids bind to the fungal elicitor cryptogein and compete with sterols.

Cryptogein is a proteinaceous elicitor of plant defense reactions which also exhibits sterol carrier properties. In this study, we report that this protein binds fatty acids. The stoichiometry of the fatty acid-cryptogein complex is 1:1. Linoleic acid and dehydroergosterol compete for the same site, but elicitin affinity is 27 times lower for fatty acid than for sterol. We show that C7 to C12 saturated and C16 to C22 unsaturated fatty acids are the best ligands. The presence of double bonds markedly increases the affinity of cryptogein for fatty acids. A comparison between elicitins and known lipid transfer proteins is discussed.