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E Blanc

Publications and source records attributed to E Blanc.

At least 19 recordsLinked to original sources
PubMed2000

A new fold in the scorpion toxin family, associated with an activity on a ryanodine-sensitive calcium channel.

We determined the structure in solution by (1)H two-dimensional NMR of Maurocalcine from the venom of Scorpio maurus. This toxin has been demonstrated to be a potent effector of ryanodyne-sensitive calcium channel from skeletal muscles. This is the first description of a scorpion toxin which folds following the Inhibitor Cystine Knot fold (ICK) already described for numerous toxic and inhibitory peptides, as well as for various protease inhibitors. Its three dimensional structure consists of a compact disulfide-bonded core from which emerge loops and the N-terminus. A double-stranded antiparallel beta-sheet comprises residues 20-23 and 30-33. A third extended strand (residues 9-11) is perpendicular to the beta-sheet. Maurocalcine structure mimics the activating segment of the dihydropyridine receptor II-III loop and is therefore potentially useful for dihydropyridine receptor/ryanodine receptor interaction studies. Proteins 2000;40:436-442.

Amino Acid Sequence↗
PubMed2000

Solution structure of BmKTX, a K+ blocker toxin from the Chinese scorpion Buthus Martensi.

BmKTX is a toxin recently purified from the venom of Buthus Martensi, which belongs to the kaliotoxin family. We have determined its solution structure by use of conventional two-dimensional NMR techniques followed by distance-geometry and energy minimization. The calculated structure is composed of a short alpha-helix (residues 14 to 20) connected by a tight turn to a two-stranded antiparallel beta-sheet (sequences 25-27 and 32-34). The beta-turn connecting these strands belongs to type I. The N-terminal segment (sequence 1 to 8) runs parallel to the beta-sheet although it cannot be considered as a third strand. Comparison of the conformation of BmKTX and toxins of the kaliotoxin family clearly demonstrates that they are highly related. Therefore, analysis of the residues belonging to the interacting surface of those toxins allows us to propose a functional map of BmKTX slightly different from the one of KTX and AgTX2, which may explain the variations in affinities of these toxins towards the Kv1.3 channels.

Amino Acid Sequence↗
PubMed2000

Extensive biopsy protocol improves the detection rate of prostate cancer.

PURPOSE: We evaluated improvement in the rate of prostate cancer detection when using an extensive biopsy protocol involving peripheral cores. MATERIALS AND METHODS: We prospectively evaluated 303 consecutive men who underwent transrectal ultrasound guided biopsy due to elevated prostate specific antigen (PSA) and/or abnormal digital rectal examination. Ten biopsies were performed, including at least 5 at the base and middle of each lobe. In addition to standard biopsy at a 45-degree angle, a more peripheral 30-degree angle biopsy was obtained. At the apex only 1 standard biopsy was done. However, when prostate volume was greater than 50 cm.3, an additional peripheral biopsy was obtained at the apex. RESULTS: The complication rate in this biopsy protocol was 1% (3 patients). Prostate cancer was detected in 118 of the 303 men (38. 9%). Overall this extensive protocol resulted in 6.6% improvement in the detection rate. Improvement was 6.5% in men with PSA 10 ng./ml. or less and 7% in those with PSA greater than 10 (not significant). CONCLUSIONS: Increasing the number of biopsy cores and improving prostate peripheral zone sampling resulted in a significant improvement in the detection of prostate cancer.

Adult↗
PubMed1999

Solution structure of potassium channel-inhibiting scorpion toxin Lq2.

Lq2 is a unique scorpion toxin. Acting from the extracellular side, Lq2 blocks the ion conduction pore in not only the voltage- and Ca2+ -activated channels, but also the inward-rectifier K+ channels. This finding argues that the three-dimensional structures of the pores in these K+ channels are similar. However, the amino acid sequences that form the external part of the pore are minimally conserved among the various classes of K+ channels. Because Lq2 can bind to all the three classes of K+ channels, we can use Lq2 as a structural probe to examine how the non-conserved pore-forming sequences are arranged in space to form similar pore structures. In the present study, we determined the three-dimensional structure of Lq2 using nuclear magnetic resonance (NMR) techniques. Lq2 consists of an alpha-helix (residues S10 to L20) and a beta-sheet, connected by an alphabeta3 loop (residues N22 to N24). The beta-sheet has two well-defined anti-parallel strands (residues G26 to M29 and residues K32 to C35), which are connected by a type I' beta-turn centered between residues N30 and K31. The N-terminal segment (residues Z1 to T8) appears to form a quasi-third strand of the beta-sheet.

Amino Acids↗
PubMed1999

Improved free R factors for cross-validation of macromolecular structure - importance for real-space refinement.

Improvements in free R cross-validation are based on changed scaling procedures and the use, in map calculation, of estimates of the validation amplitudes which are independent of the actual observed values. The deleterious effects of the omitted test data are mitigated by reduction of the test-set size, which is made possible by constraining test and working sets to share the same scaling coefficients, thereby reducing the degrees of freedom and the dependence of free R on data selection. Further improvements come with use of a modified free R factor, R freeTA. Instead of omitting the validation reflections from map calculation, their amplitudes are replaced by the average of resolution peers that is (nearly) independent of the actual cross-validation amplitudes. The improvements are relevant to model building, phase refinement by density modification and especially to real-space refinement. Although for real data at about 3 A resolution, free R factors of about 0.25 are affected little, the precision of the structure is improved by about 0.1 A. Tests with simulated data show that with good agreement between observed and calculated amplitudes (as in very high resolution studies or simulated refinement tests), free R factors can be improved by factors greater than two.

Crystallography, X-Ray↗
PubMed1999

Real-space molecular-dynamics structure refinement.

Real-space targets and molecular-dynamics search protocols have been combined to improve the convergence of macromolecular atomic refinement. This was accomplished by providing a local real-space target function for the molecular-dynamics program X-PLOR. With poor isomorphous replacement experimental phases, molecular dynamics does not improve real-space refinement. However, with high-quality anomalous diffraction phases convergence is improved at the start of refinement, and torsion-angle real-space molecular dynamics performs better than other available least-squares or maximum-likelihood methods in real or reciprocal space. It is shown that the improvements result from an optimization method that can escape local minima and from a reduction of overfitting through the implicit use of phases and through use of a local refinement in which errors in remote parts of the structure cannot be mutually compensating.

Hydroxymethylglutaryl CoA Reductases↗
PubMed1999

Critical initial real-space refinement in the structure determination of arginine kinase.

Arginine kinase (AK), a homologue of creatine kinase, catalyses the reversible transfer of a phosphoryl group between a guanidino phosphate and ADP. The family of phosphagen kinases eluded structure determination for over 25 years until an inactive form creatine kinase (CK) structure was determined [Fritz-Wolf et al. (1996). Nature (London), 381, 341-345]. The structure determination of the active-form transition-state complex was non-trivial, owing to the distant relatedness and domain reorientation of AK compared with CK. Phases from a molecular-replacement solution of the large domain, supplemented by single isomorphous replacement and inter-crystal averaging, did not reveal interpretable electron density for the small domain. Reciprocal-space refinement of the initial model (Rfree = 0.54) by any of the commonly used methods, including post facto application of maximum-likelihood methods, led to overfitting without significant improvement of the partial initial model. By contrast, in the local real-space refinements which proved successful, the interdependence of atoms is limited to immediate neighbors, and atomic positions are not influenced by errors or omissions in remote parts of the structure. Modest improvement was possible without overfitting, and this was critical to the calculation of improved phases. Phases were refined and extended from 4.0 to 2.5 A resolution by Fourier inversion of omit maps, combination with isomorphous replacement phases and averaging between crystal forms, after several batches of real- and reciprocal-space atomic refinement. The final structure refinement, against a 1.86 A cryo data set yielded a high-quality model with R = 0.196 and Rfree = 0.224.

Arginine Kinase↗
PubMed1999

Diagnostic value of ten systematic TRUS-guided prostate biopsies.

OBJECTIVE: To evaluate the improvement in the rate of detection of prostate cancer using an extensive protocol involving ten transrectal biopsies. METHODS: A total of 162 patients submitted to transrectal ultrasound-guided biopsy for elevated prostate-specific antigen (PSA) and/or abnormality on digital rectal examination were studied consecutively and prospectively. Five biopsies were performed in each lobe: between the three standard biopsies on each side, two additional biopsy specimens were taken in the same plane and at the same 45 degrees angle. RESULTS: The complication rate with the ten-biopsy protocol was 1.85%. Prostate cancer was detected in 40.1% of the patients. In the overall series, the percentage of diagnostic improvement brought about by this ten-biopsy protocol was +3.1%. The percentage improvement was greatest (+4.9%) in patients with PSA </=10 ng/ml. CONCLUSION: Increasing the number of biopsy cores without altering the angle of biopsy and/or the zone sampled does not lead to a significant improvement in the detection of prostate cancer.

Adult↗
PubMed1999

[Value of the use of a questionnaire in the evaluation of incontinence surgery].

OBJECTIVE: To assess the contribution of a questionnaire in the evaluation of patients operated for urinary stress incontinence. PATIENTS AND METHODS: From 1988 to 1996, 42 patients underwent bladder neck suspension for urinary stress incontinence. Only 7 patients had no history of pelvic surgery, 24 patients presented pure urinary stress incontinence and 19 presented mixed incontinence, 37 patients were evaluable with a mean follow-up of 2 years. The evaluation consisted of two aspects: clinical assessment, based on review of the case files, showing that 77% of patients were continent, and a questionnaire (non-validated translation of "Q7" and "UD16"). This questionnaire evaluated quality of life (7 items) and symptoms. Each item was scored from 0 to 3. RESULTS: To interpret the results we divided the three main scores into three intervals, the first third corresponding to good results. The percentage of good results for the overall score, quality of life score and symptom score was 62%, 73% and 46% respectively. The most discriminative questions of the questionnaire can be used to analyse urine leaks related to effort, urgency episodes of incontinence. A score of 0 or 1 was considered to be a good result. The percentage of good results for these three questions was therefore 72%, 60% and 64%, respectively. CONCLUSION: The cure rate therefore varies as a function of the criteria selected: the questionnaire allows the urologist to more accurately assess the patient's postoperative state.

Adult↗
PubMed1998

Solution structure of two new toxins from the venom of the Chinese scorpion Buthus martensi Karsch blockers of potassium channels.

The solution structure of BmTX2 purified from the venom of the Chinese Buthid Buthus martensi has been determined by 2D NMR spectroscopy techniques which led to the description of its 3D conformation. The structure consists of a triple-stranded beta-sheet connected to a helical structure. This helix encompasses 10 residues, from 11 to 20, begins with a turn of 310 helix, and ends with an alpha helix. The three strands of beta sheet comprise residues 2-6, with a bulge covering residues 4 and 5, 26-29, and 32-35, with a type I' beta turn centered on residues 30-31. We also characterized the solution structure of BmTX1. The two toxins which are potent blockers of both large-conductance calcium-activated potassium channels (BKCa channels) and voltage-gated potassium channels (Kv1. 3) are highly superimposable and possess the same structural characteristics. Analysis of these structures allows us to hypothesize that, besides the main surface of interaction described by the functional map of charybdotoxin, one can expect that the binding of scorpion toxins on BKCa channels may involve residues on the edge of this surface.

Amino Acid Sequence↗
PubMed1998

Transition state structure of arginine kinase: implications for catalysis of bimolecular reactions.

Arginine kinase belongs to the family of enzymes, including creatine kinase, that catalyze the buffering of ATP in cells with fluctuating energy requirements and that has been a paradigm for classical enzymological studies. The 1.86-A resolution structure of its transition-state analog complex, reported here, reveals its active site and offers direct evidence for the importance of precise substrate alignment in the catalysis of bimolecular reactions, in contrast to the unimolecular reactions studied previously. In the transition-state analog complex studied here, a nitrate mimics the planar gamma-phosphoryl during associative in-line transfer between ATP and arginine. The active site is unperturbed, and the reactants are not constrained covalently as in a bisubstrate complex, so it is possible to measure how precisely they are pre-aligned by the enzyme. Alignment is exquisite. Entropic effects may contribute to catalysis, but the lone-pair orbitals are also aligned close enough to their optimal trajectories for orbital steering to be a factor during nucleophilic attack. The structure suggests that polarization, strain toward the transition state, and acid-base catalysis also contribute, but, in contrast to unimolecular enzyme reactions, their role appears to be secondary to substrate alignment in this bimolecular reaction.

Animals↗
PubMed1998

Determination of the relative precision of atoms in a macromolecular structure.

Several real-space indices and temperature factors are compared with respect to their correlation with atomic positional error and their ability to indicate atoms and residues with the worst of subtle errors. The best index, rED, is a correlation coefficient between model and map electron densities, similar to one proposed earlier, but incorporating two improvements. Firstly, resolution is accounted for explicitly by calculating the model electron density by Fourier transformation of resolution-truncated scattering factors. Secondly, the deviation between model and map electron densities is assigned to neighboring atoms according to their contribution to the electron density of each grid point. With maps of various qualities, rED is the single index with best correlation to atomic error with grouped or individual atoms, and it is the most reliable indicator of poor residues. With poorer omit maps, imprecision of individual atoms is best diagnosed by a combination of low rED or high B factor. With the improved methods, 60-70% of the least precise atoms can detected in a fully refined structure. Similarly, 40-80% of the least precise atoms of an unrefined model can be detected by comparison with an isomorphous replacement map. This is useful in assessing and improving the quality of a model, but not sufficient to confidently validate all atoms of a structure at sub-atomic resolution.

Electrons↗
PubMed1998

[Preliminary results of the treatment of 44 patients with localized cancer of the prostate using transrectal focused ultrasound].

OBJECTIVE: To evaluate the efficacy and morbidity of treatment of localized prostatic cancer using transrectal high-intensity focused ultrasound. MATERIAL AND METHODS: 44 patients (mean age: 72 years) with clinical stage T1 (20) or T2 (24) prostatic cancer, not eligible for radical prostatectomy, were treated by two different prototypes of the Ablatherm (Technomed Medical System). The second prototype includes several safety devices designed to reduce morbidity. 99 sessions were performed, i.e. an average of 2.25 sessions per patient: the prostate was treated in one session (5 patients), 2 sessions (26 patients), 3 sessions (10 patients), or 4 sessions (3 patients), usually under spinal anaesthesia. The mean PSA was 9.92 ng/ml and the mean prostatic weight was 37 g. RESULTS: Complications occurred in 10 (50%) of the first 20 patients treated (1993-1995): 2 recto-urethral fistulas, 2 asymptomatic rectal burns, 2 cases of stable urinary retention, 1 case of severe incontinence, 3 cases of bladder neck sclerosis, and 1 febrile urinary tract infection. In the 24 patients treated subsequently (1995-1997), complications occurred in only 4 patients (16%): 1 case of stable urinary retention, 1 febrile urinary tract infection, 1 case of bladder neck sclerosis, 1 case of stress incontinence. A complete response (repeated negative follow-up biopsies) was obtained in 27 patients (61%). The mean post-treatment PSA level in patients of this group was 1.09 +/- 1.06 and remained stable (mean follow-up: 346 days; range: 60-1250). A residual cancer was detected in 17 patients (39%). 8 patients were considered to be failures (mean post-treatment PSA: 4.8 ng/ml) and received adjuvant treatment (hormonal: 3; external radiotherapy: 5). Complementary treatment by focused ultrasound will be performed in another 9 patients, if their PSA rises above 3 ng/ml (patients of this group are symptomatic with a mean post-treatment PSA of 1.6 ng/ml). CONCLUSION: The morbidity of treatment by transrectal focused ultrasound has now been reduced. These results suggest that this new treatment can constitute a useful option for certain patients with localized cancer, not suitable for radical prostatectomy.

Adenocarcinoma↗
PubMed1998

[Diagnostic value of 10 systematized and ultrasound-guided transrectal prostatic biopsies].

OBJECTIVE: To study the improvement of prostatic cancer detection provided by ten strictly systematized transrectal prostatic biopsies. MATERIAL AND METHODS: This prospective study was conducted in 162 patients submitted to a series of 10 ultrasound-guided transrectal prostatic biopsies due to the presence of elevated PSA and/or an abnormality on digital rectal examination. Five biopsies were taken from different sites in each lobe: 2 biopsies were inserted between the 3 biopsies usually performed in the standard protocol, while maintaining the angle of entry of the needle recommended in this protocol. RESULTS: The complication rate was 1.85% with the 10-biopsy technique. Prostatic cancer was detected in 40.1% of the 162 patients. The percentage diagnostic improvement provided by the 10-biopsy protocol in the overall patient population was +3.1% compared to the standard protocol. The highest percentage diagnostic improvement was observed in the group of patients with PSA < or = 10 ng/ml (+4.9%) and in the group of patients with an ultrasound prostatic volume < or = 40 cc (+4%). CONCLUSION: The most marked diagnostic improvement appears to be related more to sampling of prostatic zones presenting a statistically increased risk of cancer (peripheral zone) than to the increased number of prostatic biopsies performed.

Adenocarcinoma↗
PubMed1997

1H-NMR-derived secondary structure and overall fold of a natural anatoxin from the scorpion Androctonus australis hector.

The venom of the scorpion Androctonus australis hector contains several protein neurotoxins of which structure and structure/activity relationships have been extensively studied. It also contains polypeptides such as Aah STR1, which are not toxic, while having highly similar sequences to fully active toxins. We have determined the solution structure of Aah STR1 by use of conventional two-dimensional NMR techniques followed by distance-geometry and energy minimization. We have demonstrated that, despite its lack of toxicity, Aah STR1 is structurally highly related to anti-mammal scorpion toxins specific for Na+ channels. The calculated structure is composed of a short alpha-helix (residues 26-33) connected by a tight turn to a three-stranded antiparallel beta-sheet (sequences 3-6, 38-41 and 44-48). This beta-sheet is right-handed twisted as usual for such secondary structures. The beta-turn connecting the strands 38-41 and 44-48 belongs to type II'. The overall fold of Aah STR1 is typical of beta-type scorpion toxins. This is, however, the first example of such a fold in Old World scorpion toxins. Either the absence of a basic residue in position 63 or the high mobility of loops, compared to active beta-type neurotoxins, may explain the lack of activity of this protein.

Amino Acid Sequence↗
PubMed1997

Solution structure of TsKapa, a charybdotoxin-like scorpion toxin from Tityus serrulatus with high affinity for apamin-sensitive Ca(2+)-activated K+ channels.

TsKapa (TsK), purified from the Buthidae Tityus serrulatus is a very high potent ligand for small-conductance apamin-sensitive calcium-activated potassium channels (SK). It is able to efficiently compete with apamin for binding on this channel (K0.5 = 0.3 nM) [Legros, C. et al., FEBS Lett. 390:81-84, 1996]. The solution structure of TsK has been determined by 2D-NMR techniques, which led to the full description of its 3D conformation: a short alpha helix from residues 14 to 20 and a three-stranded antiparallel beta sheet (residues 2-3, 27-29, and 32-34). The interaction of TsK with the SK potassium channel has been modeled according to the charge anisotropy of the ligand. The resulting dipole moment orientates TsK so that it presents toward the receptor, a surface, mainly basic, encompassing residues K18 and K19 on one side and R9 and Y8 on the other. Despite its three-dimensional structure that is related with scorpion toxins active on voltage-gated potassium channels such as charybdotoxin, the pharmacological activity and specificity of TsK is related with shorter scorpion toxins (i.e., possessing an only two-stranded beta sheet) such as scyllatoxin (also named leiurotoxin I) or P05.

Amino Acid Sequence↗