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B Scharf

Publications and source records attributed to B Scharf.

At least 37 records · Page 2Linked to original sources
PubMed1994

Blue halorhodopsin from Natronobacterium pharaonis: wavelength regulation by anions.

Halorhodopsin, the chloride pump from Natronobacterium pharaonis (pharaonis hR), was isolated under conditions of low ionic strength. The quotient between the optical densities of pharaonis hR in 4 M NaCl at 280 and 577 nm amounts to 1.1, indicating a high purity of the protein and integrity of the chromophore. Gel filtration chromatography of the purified pharaonos hR at neutral pH and in the absence of inorganic salts leads to a shift to the absorption maximum to 600 nm (pharaonis hRblue). The purple color can be regained by the addition of anions such as chloride, iodide, azide, nitrate, and also fluoride. The absorption maxima are dependent on the nature of the anion and the pH. At pH 7, sulfate does not influence the 600-nm absorption, while at pH 4.5 the absorption maximum is shifted to 581 nm. The blue form of halorhodopsin (pharaonis hRblue) was titrated with different sodium salts, such as chloride, azide, and nitrate. The half-maximal binding is in the millimolar range, with Br < Cl < NO3 < N3 < BrO3. Deprotonation of the Schiff base can be accomplished by treatment of pharaonis hR or pharaonis hRblue with base. The apparent pK of the Schiff base in pharaonis hRblue was determined to be 8.5. The pK shifted to 8.0 in the presence of 150 mM Na2SO4, whereas the pK of pharaonis hR in 150 mM NaCl is 9.6. In the photocycle of the chloride- and nitrate-containing pharaonis hR, a species similar to hR520 could be detected. Pharaonis hR reconstituted with azide shows photochemical reactions similar to the photocycle of bacteriorhodopsin.(ABSTRACT TRUNCATED AT 250 WORDS)

Anions↗
PubMed1994

The primary structure of halocyanin, an archaeal blue copper protein, predicts a lipid anchor for membrane fixation.

Halocyanin, a small blue copper protein, was isolated from the haloalkaliphilic archaeon Natronobacterium pharaonis. The NH2 terminus was not accessible to Edman degradation. About 70% of the amino acid sequence was determined by protein sequence analysis. The sequence information of two peptides was used for cloning and sequencing the halocyanin gene (hcy). The open reading frame codes for 489 base pairs, which account for a protein with 163 amino acids and a molecular mass of 17,223 Da. The discrepancy between this value and the molecular mass of 15,456 +/- 1.5 Da for the copper-free protein determined by electrospray mass spectrometry can be explained by a post-translational processing of the gene product. The NH2-terminal sequence of the open reading frame contains a motif that is characteristic for prokaryotic lipoproteins. Assuming a similar processing for halocyanin, Cys at position 25 of the primary transcript would be modified by a diphytanyl (glycerol)diether. Subsequently, the precursor is cleaved by a signal peptidase II-like protease and then acetylated at its NH2-terminal alpha-amino group. These modifications would yield a protein with a calculated molecular mass of 15,456 Da. A comparison of the primary structure of halocyanin with a number of other blue copper proteins places it into the plastocyanin-related group.

Amino Acid Sequence↗
PubMed1994

Phototactic behaviour of the archaebacterial Natronobacterium pharaonis.

Natronobacterium pharaonis can react tactically to photo- and chemostimuli. It moves by rotation of a flagellar bundle which is monopolarly inserted. Under sufficient oxygen supply the photophobic response of N. pharaonis has been measured. The resulting action spectrum matches the absorption spectrum of the purified retinylidene protein psR-II. Retical synthesis could be inhibited by nicotine. Cells grown in the presence of nicotine show a strongly reduced photoresponse, which could be restored by addition of retinal. These data identify psR-II as the receptor for negative phototaxis.

Chemotaxis↗
PubMed1994

On the role of the olivocochlear bundle in hearing: a case study.

A young patient with normal pure-tone thresholds in both ears underwent a unilateral vestibular neurotomy in January 1992 to relieve severe vertigo ascribed to Ménière's disease. Evidence is provided that the whole vestibular nerve including the olivocochlear bundle (OCB) was sectioned. Just prior to the surgery, the patient was examined in several psychoacoustic tests involving mainly signal detection and selective attention. Over the next 20 months, he was reexamined in those same tests. The patient's ability to detect expected tones in the quiet (including audiograms) or in noise was the same as before the surgery. The one change was a marked improvement in the detection of unexpected signals in noise, which appears to reflect impaired selective attention. During those 20 months, new tests were also performed on discrimination, loudness, pitch, lateralization, and temporary threshold shift. On these tests, the only differences between the operated and unoperated ears concerned binaural diplacusis and loudness adaptation close to threshold, but these differences may well have been present prior to the surgery. Except with respect to what is probably selective attention, we uncovered no other clear role for the OCB in hearing. This outcome agrees with limited measurements on other patients, with their subjective reports, and with a number of published neurophysiological observations.

Acoustic Stimulation↗
PubMed1993

Analysis of the redox reaction of an archaebacterial copper protein, halocyanin, by electrochemistry and FTIR difference spectroscopy.

Halocyanin is a recently discovered archaebacterial copper protein classified as "type I" small blue copper protein (Scharf, B., Ph.D. Thesis, University of Bochum, Germany). Its redox properties were investigated by a combination of protein electrochemical and spectroscopic techniques. Using electrochemical reactions in an ultrathin-layer electrochemical cell developed for UV/vis and IR spectroscopy, halocyanin could be quantitatively and reversibly oxidized and reduced. The titration of the absorption band at 600 nm can be perfectly described by a Nernst curve with n = 1 electron transferred; a quantitative fit yields a midpoint potential, Em, of 183 mV (vs SHE) at a pH of 7.3. The midpoint potential falls constantly from +333 mV at pH 4 to +119 mV at pH 10, with three regions around pH 4.5, 6.5, and 8.5 where the pH dependence is ca. -60 mV/pH unit, indicating the uptake of a proton with the reduction. By analogy with other small type I copper proteins, the three pK values suggested by the pH dependency of Em might be associated with three histidines which interact with the redox site. Electrochemically induced reduced-minus-oxidized Fourier transform infrared difference spectra in the 1800-1000 cm-1 range at neutral pH show a number of strong difference bands between ca. 1700 and 1600 cm-1 as well as smaller difference structures between 1600 and 1200 cm-1. The maximum amplitude of the difference bands--only ca. 1% of the amide-I absorption at ca. 1639 cm-1--indicates that only small protein rearrangements occur upon the redox transition.(ABSTRACT TRUNCATED AT 250 WORDS)

Archaea↗
PubMed1993

Halocyanin, an archaebacterial blue copper protein (type I) from Natronobacterium pharaonis.

A small blue copper protein with a molecular mass of about 15.5 kDa has been isolated from the haloalkaliphilic Natronobacterium pharaonis. This protein, which was named halocyanin, contains one Cu2+. The circular dichroism spectrum in the region of the absorption of the peptide bond reveals mainly beta-structural elements. The absorption spectrum in the visible range has three maxima with the main band at 600 nm. The circular dichroism spectrum in this region is characterized by four extrema with maxima at 413 and 590 nm and minima at 477 nm as well as 721 nm. The electron paramagnetic resonance spectrum displays a hyperfine structure which shows a close resemblance to those of plastocyanin. The cumulative spectral data agree well with those of other small blue copper proteins with axial symmetry of the Cu ligands. In analogy, one can deduce that halocyanin may adopt a type I copper binding site with two His, one Met, and one Cys as probable ligands. This conclusion is confirmed by the C-terminal sequence which contains, homologous to other known sequences of type I copper proteins, three of the four copper ligands. Halocyanin can only be removed from the membrane by mild treatment with detergents. This observation indicates that halocyanin is a peripheral membrane protein which serves as a mobile electron carrier. The presence of type I blue copper proteins is archaebacteria might lead to further insights into their phylogenetic origin.

Amino Acid Sequence↗
PubMed1993

Loudness adaptation at high frequencies.

Loudness adaptation was measured for pure tones at 4, 12, 14, and 16 kHz. In three experiments, a total of 87 young listeners judged--by the method of successive magnitude estimation--the loudness of the tones over a 6-min exposure period. Thresholds were measured by an adaptive 2IFC procedure. Although earlier measurements had shown that adaptation near threshold increases with frequency, these new data reveal that the increase is especially marked at higher sensation levels. Thus, at 40 dB SL, over a 6-min period loudness declined by 18% at 4 kHz and by 94% at 16 kHz. Moreover, the 4-kHz tone remained audible for all listeners throughout the 6-min exposure period whereas the 16-kHz tone became inaudible for two-thirds of them by the end of the exposure period. Listeners with relatively low thresholds (< 50 dB SPL) at 16 kHz showed much less adaptation at 14 kHz than at 16 kHz and much less than listeners with relatively high thresholds (> 50 dB SPL) at 16 kHz; this latter group showed strong adaptation at 14 kHz. The marked loudness adaptation of steady tones at very high frequencies and relatively high sensation levels is associated with a restricted spread of excitation in the auditory system resulting from the steep rise of the threshold curve at the highest audible frequencies.

Acoustic Stimulation↗
PubMed1993

Loudness adaptation in children.

Simple loudness adaptation was measured for a steady tone presented alone at 10 dB SL; contralaterally induced adaptation was measured for a steady tone in one ear accompanied by an intermittent tone in the contralateral ear; ipsilaterally induced adaptation was measured for a tone increased intermittently by 15 dB. The method of successive magnitude estimation revealed no differences between 12 adults and 36 children from 9 to 14 years of age in the amounts of adaptation over a 3-min exposure. A second set of experiments with a new group of 20 adults and 20 children used a Békésy tracking procedure to reach similar results. Unlike Kärjä [Acta Oto-Laryngol 1968; (suppl)241:1-56], who found significant adaptation in only 5 of 29 children and much adaptation in most adults, we found considerable adaptation in children as well as in adults.

Acoustic Stimulation↗
PubMed1992

Chromophore of sensory rhodopsin II from Halobacterium halobium.

The photoreceptor sensory rhodopsin II (sR-II) was enriched 120-fold from cell membranes of Halobacterium halobium. The final preparation yields sR-II with a specific content of 3 nmol of sR-II/mg of protein. The spectroscopic measurements were performed on the enriched photoreceptor solubilized in digitonin. In the absolute absorption spectrum of the partially purified receptor, the main peak in the visible range corresponded to sR-II with a maximum at 488 nm. Cytochromes contributed to the spectrum only in a minor band at 415 nm. The extinction coefficient of sR-II was estimated from difference spectra during bleaching with hydroxylamine to be 48,000 M-1 cm-1. The reduced chromophore displayed a pronounced fine structure which is due to the coplanarity of the retinyl residue. The isomeric composition of the chromophore from the enriched photoreceptor was determined in retinal extracts in HPLC. The dark-adapted sR-II contains 80% all-trans- and 20% 13-cis-retinal. After illumination, the ratio changed to 1:1, indicating a trans-cis isomerization during the photocycle of sR-II.

Archaeal Proteins↗
PubMed1992

Biochemical and photochemical properties of the photophobic receptors from Halobacterium halobium and Natronobacterium pharaonis.

The phototaxis of Halobacterium halobium is initiated by two photoreceptors, the sensory rhodopsins sR-I and sR-II. An sR-II-like pigment has also been described in Natronobacterium pharaonis. In this work it was shown that N. pharaonis cells are repelled by light with a wavelength of 500 nm. A further comparison of membrane preparations from H. halobium (mutant D1) containing only sR-II and from N. pharaonis [strain SP1(28)] with a chromophoric protein (psR-II) resembling sR-II revealed substantial similarities. The biochemical and photochemical properties of the pigments are quite similar, with psR-II being more stable to external conditions such as pH and ionic strength of the buffer. Both pigments are bleached by low concentrations of hydroxylamine and can be reconstituted by the addition of all-trans-retinal. The absorption spectrum of psR-II is quite similar to sR-II including the shoulder on the short-wavelength side. After light excitation sR-II and psR-II undergo photocycles with at least three intermediates. The earliest intermediate has an absorption maximum above 520 nm and decays to a species which has a characteristic absorption (approximately 380 nm) of a deprotonated Schiff base. The final step is the regeneration of the original ground state via a red-shifted intermediate absorbing around 540 nm. From this cumulative evidence it can be concluded that, not only sR-II, but also the pigment from N. pharaonis is a photophobic photoreceptor.

Archaea↗
PubMed1991

Effective attenuation of signals in noise under focused attention.

When attending to a tone at a given frequency, listeners are most sensitive to that tone and others within a restricted band of frequencies surrounding it. This region of enhanced sensitivity defines the attention band that was measured in two experiments using a modified version of the probe-signal method of Greenberg and Larkin [J. Acoust. Soc. Am. 44, 1513-1523 (1968)]. Experiment 1 showed that at five center frequencies, from 0.25 to 4.0 kHz, the shape of the attention band resembles that of the auditory filter as inferred from notched-noise masking experiments by other investigators. The width of the attention band is close to the critical band at higher frequencies, but only half as wide at 0.25 and 0.5 kHz. Experiment 2 produced psychometric functions for unattended probe tones at least 0.23 kHz away from a fully attended, 1-kHz target tone. From these functions, the effective attenuation, measured as the threshold difference between the 1-kHz target and the probes, was estimated to be 7 dB; the amount of attenuation appeared to be about the same regardless of how far the probe frequency was from the attended band. One interpretation of these results is that bands centered on the unattended tones contribute to the decision process with some small but measurable weight and are not entirely ignored.

Acoustics↗
PubMed1990

The loudness of sounds that increase and decrease continuously in level.

A sound at a low level is heard as much softer after having decreased continuously from higher levels than if presented after a period of silence at that same low level. Canévet [Acustica 61, 256-264 (1986)] demonstrated this phenomenon for a tone that (1) decreased from 65 to 20 dB in 180 s; he also presented a tone that (2) increased from 20 dB, or (3) was presented as pairs of bursts at various levels in random order. Below about 40 dB, loudness changed most rapidly in the decreasing condition so that, at 20 dB, the tone was judged ten times softer than in conditions (2) and (3). In the present experiments, magnitude estimation was used to examine the possible role of judgmental biases and adaptation in this rapid loudness decline, which we call decruitment. Results show that decruitment did not come about because subjects made many successive loudness judgments; loudness declined as much when a tone was judged only twice, at the beginning and end of its 180-s decrease. In contrast, interrupting the decreasing tone so that it was heard only at 70 dB and 160 s later at 30 dB greatly diminished the decruitment. Similarly, pairs of 500-ms tone bursts presented at successively lower levels instead of continously decreasing did not show decruitment, suggesting that sequential biases are irrelevant. The likely cause of decruitment is sensory adaptation.

Attention↗
PubMed1987

On the relation between the growth of loudness and the discrimination of intensity for pure tones.

The intensity jnd is often assumed to depend on the slope of the loudness function. One way to test this assumption is to measure the jnd for a sound that falls on distinctly different loudness functions. Two such functions were generated by presenting a 1000-Hz tone in narrow-band noise (925-1080 Hz) set at 70 dB SPL and in wideband noise (75-9600 Hz) set at 80 dB SPL. Over a range from near threshold to about 75 dB SPL, the loudness function for the tone is much steeper in the narrow-band noise than in the wideband noise. At 72 dB SPL, where the two loudness curves cross, the tone's jnd was measured in each noise by a block up-down two-interval forced-choice procedure. Despite the differences in slope (and in sensation level), the jnd (delta I/I) is nearly the same in the two noises, 0.22 in narrow-band noise and 0.20 in wideband noise. The mean value of 0.21 is close to the value of 0.25 interpolated from Jesteadt et al. [J. Acoust. Soc. Am. 61, 169-176 (1977)] for a 1000-Hz tone that had the same loudness in quiet as did our 72-dB tone in noise, but lay on a loudness function with a much lower slope. These and other data demonstrate that intensity discrimination for pure tones is unrelated to the slope of the loudness function.

Acoustic Stimulation↗
PubMed1986

Loudness reduction and adaptation induced by a contralateral tone.

An intermittent tone in one ear may induce a large decline in the loudness of a continuous tone in the contralateral ear [Botte et al., J. Acoust. Soc. Am. 72, 727-739 (1982)]. To uncover the basis for this induced loudness adaptation, the method of successive magnitude estimations was used to measure the loudness of a test tone in one ear during and after a single presentation of a brief inducer tone in the contralateral ear. Duration and frequency of the inducer were varied. The frequency of the test tone was set at 500, 1000, or 3000 Hz. Both inducer and test tones were at 60 dB SPL. When the inducer lasted 5 s or more and was at the same frequency as the test tone, the loudness of the test tone was reduced by 80% to 100% while the inducer was on. As the inducer frequency moved away from the test tone, the loudness reduction declined gradually except for a more marked drop at the point where the frequency separation exceeded the critical bandwidth. Loudness remained depressed after the inducer went off. Additional measurements showed that the amount of loudness reduction corresponded closely to the measured movement of the inducer's sound image away from the center of the listener's head (decentralization).

Acoustic Stimulation↗
PubMed1986

[Auditory localization and speech perception in noise. Preliminary study concerning 5 cases of perceptual deafness].

The experiments reported here examine the psychoacoustical bases for poor speech perception in noise by persons with sensorineural impairment. Two major hypotheses are tested. First, because persons with cochlear impairment are less able than normal-hearing persons to separate out incoming signals on the basis of spectral differences (a deficit referred to as reduced frequency selectivity), they are less able to localize one sound in the presence of other sounds. Second, this reduced localization ability makes it difficult for the hearing-impaired person to take advantage of the spatial separation of a target speech source and other interfering sources. Such separation is common in real environments and facilitates speech perception by normal-hearing persons. Tests of these hypotheses are conducted by means of detailed psychoacoustical measures of frequency selectivity, of localization and speech perception under masking.

Adolescent↗
PubMed1985

Simple and induced loudness adaptation.

Simple loudness adaptation is the decrease in loudness that takes place when a continuous sound is presented alone for a period of time. Simple adaptation normally occurs only when a sound is soft to begin with, no more than 30 dB above threshold; except for some persons with a retrocochlear lesion, sounds above 30 dB SL do not diminish in loudness over time. However, adaptation can be induced in at least two ways: (1) A steady sound to one ear, presented together with an intermittent sound to the contralateral ear, decreases in loudness by 50-60% within 3 min. (2) An otherwise steady sound that is intermittently increased in level by at least 5 dB becomes softer during its weaker periods. When, for example, a 40-dB tone is increased every 20 s to 60 dB for 15 s, its loudness decreases by about 50% within 3 min. We report measurements of both simple and induced adaptation on 10 persons listening to a 1 000-Hz tone via earphones or from a loudspeaker. The results provide an overview of both types of adaptation. They also permitted a correlational analysis that reveals some of the similarities and differences between the two kinds of adaptation.

Adaptation, Physiological↗
PubMed1984

Lateralization and frequency selectivity in normal and impaired hearing.

The onset-time difference delta T required to lateralize a 30-ms bifrequency tone burst toward the leading ear was measured as a function of the frequency difference delta F between the tone in the left ear and the tone in the right ear. At center frequencies of 0.5 and 4 kHz, four normal listeners tested at 80 and 100 dB SPL had delta Ts that were relatively constant at subcritical delta Fs, but increased at delta Fs wider than a critical band. At 1 kHz, delta T increased with delta F even at subcritical delta Fs. Ten listeners with cochlear impairments were tested at 100 dB SPL. Seven had normal delta Ts at 4 kHz, despite hearing losses between 50 and 70 dB. At 0.5 and 1 kHz, mildly impaired listeners had nearly normal lateralization functions, whereas more severely imparied listeners had very large delta Ts and no frequency selectivity. These and other findings indicate that listeners even with moderate to severe hearing losses can lateralize normally on the basis of interaural differences in onset envelope, but not on the basis of temporal differences in the fine structure.

Adolescent↗