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J F Koretz

Publications and source records attributed to J F Koretz.

At least 37 records · Page 2Linked to original sources
PubMed1990

Hydrostatic pressure studies of native and synthetic thick filaments: II. Native thick filaments from rabbit skeletal muscle.

Native thick filaments isolated from freshly prepared rabbit psoas muscle were found to be resistant to pressure-induced dissociation. With increasing pressure application and release, a bimodal distribution of filament lengths was observed. The shorter filament length is associated with filament breakage at the center of the bare zone, while the longer length is associated with relatively intact filaments. Intact filaments and filament halves decrease in length by no more than 20% after exposure to and release of 14,000 psi. Bimodal distributions were not observed in equivalent experiments performed on filaments isolated from muscle glycerinated and stored at -20 degrees C for 6 months. Instead, filament dissociation proceeds linearly as a function of increasing pressure. Filaments prepared from muscle glycerinated and stored for 2 and 4 months exhibited pressure-induced behavior intermediate between the filaments prepared from fresh muscle and filaments prepared from muscle stored for 6 months. Since there appears to be no difference in the protein profiles of the various muscle samples, it is possible that stabilization of the native thick filament against hydrostatic pressure arises from trapped ions that are leached out over time.

Actin Cytoskeleton↗
PubMed1989

The effect of phosphorylation on the structure of alpha-crystallin.

Homopolymers were constructed from the highly purified phosphorylated (A1 and B1) and non-phosphorylated (A2 and B2) polypeptides of alpha-crystallin. These were examined using electron microscopy, light scattering, fluorescence spectroscopy and sulphydryl probing to determine the effects of phosphorylation on the structure of alpha-crystallin. Each reconstituted aggregate consisted of uniform particles with circular cross-sections and diameters of 9.3-9.5 nm. Some of these were associated in chain-like structures. The molecular mass of the homopolymers varied from 360-507 kDa; for a single particle, it was estimated to be 216 +/- 10 kDa. Tryptophan residues in the alpha B2 homopolymers were more accessible to the solvent and to quenchers than those in alpha A2. Phosphorylation increased this accessibility in the alpha A homopolymer but decreased it in alpha B. This decrease could be attributed to phosphorylation of the serine adjacent to tryptophan 60 in the B chain. The kinetics of the reaction with DTNB indicated that the single cysteine in the A chain was buried in the alpha A2 homopolymer (k1' = 0.013 min-1) but more accessible in alpha A1 (k1' = 0.046 min-1). It was concluded that phosphorylation significantly alters the conformation of the alpha A subunits but probably has little effect on the B subunits. The alterations do not affect the ability of the subunits to associate into alpha-crystallin-like particles.

Animals↗
PubMed1989

Hydrostatic pressure studies of native and synthetic thick filaments: in vitro myosin aggregates at pH 7.0 with and without C-protein.

Column-purified myosin at pH 7.0 will reproducibly aggregate into filaments of known average length and structure when dialyzed against a low ionic strength medium under controlled conditions. When exposed to increased hydrostatic pressure, followed by quick return to atmospheric pressure, the original filaments shorten linearly with increasing pressure; in addition, a second population of filaments is seen, presumably the result of reaggregation of myosin after release of pressure. This second population is about 0.5 microns long, bipolar, and about half the diameter of the original filaments. The number of these filaments, but not their physical characteristics, is a function of the shortening of the original filament population. Both the remnants of the original population and the new aggregates, once formed, are stable over time and at room temperature. The addition of C-protein to myosin solutions before filament preparation results in a filament population of slightly shorter length. When these filaments are exposed to increased hydrostatic pressure, they are more resistant to disaggregation than myosin filaments without C-protein. However, like the filaments prepared in the absence of C-protein, a second population of shorter, thinner filaments is visible after exposure to pressure.

Animals↗
PubMed1989

Accommodation and presbyopia in the human eye--aging of the anterior segment.

Ocular biometric parameters and accommodative amplitude were measured by various techniques in 100 normal emmetropic human subjects age 18-70 yr. Anterior chamber depth decreased and lens thickness increased linearly over the entire age group. Accommodative amplitude declined linearly until a stable nadir was reached at about age 50 yr. The respective slopes and intercepts of the age-dependent decline in anterior chamber depth were essentially the same for measurements made independently by optical pachmetry, A-scan ultrasonography, and slit-lamp Scheimpflug photography. The age-dependent increase in lens thickness differed in slope and intercept for measurements made by photography and ultrasonography if the generally accepted lenticular sound velocity was assumed for all subjects. However, if putative lenticular sound velocity was adjusted for age, the relationships given by the two techniques were essentially identical. Total anterior segment length (defined as the distance between the anterior corneal and posterior lens surfaces), vitreous cavity length (distance between the posterior lens and anterior retinal surfaces), and total globe length were all independent of age. This constellation of findings indicates that the human lens grows throughout adult life while the globe does not, that thickening of the lens completely accounts for shallowing of the anterior chamber with age, but that the posterior surface of the lens remains fixed in position relative to the cornea and retina.

Accommodation, Ocular↗
PubMed1989

The effect of temperature on the renaturation of alpha-crystallin.

The effects of variations in temperature and protein concentration on the renaturation of bovine alpha-crystallin have been examined using gel permeation chromatography, sedimentation analysis, fluorescence spectroscopy and electron microscopy. High protein concentration (3-53 mg/ml) were found to generate heterogenous populations of aggregates. It was concluded that concentrations above 3 mg/ml were inappropriate for renaturation of alpha-crystallin. Aggregates with molecular masses gradually increasing from 461,000 to 695,000 Da were produced with increasing temperature over the range 6-39 degrees C. Electron microscopy demonstrated that the reaggregates were composed predominantly of particles with circular cross-sections and mean diameters of 13-14 nm. As the renaturation temperature increased, increasing amounts of sheet-like structures were observed. Tryptophan accessibility to acrylamide quenching decreased in these aggregates as the size increased. These observations are consistent with the concept that there is no unique quaternary structure, or set of structures, for alpha-crystallin but that the protein can exist in a variety of forms containing different numbers of subunits.

Animals↗
PubMed1988

Slit-lamp studies of the rhesus monkey eye: III. The zones of discontinuity.

The rhesus monkey lens exhibits two zones of discontinuity, one anteriorly and one posteriorly. They are first discernible by slit-lamp photography as performed in this study at around age 7 years in iridectomized eyes, and become more distinct with increasing age. Their thickness and distance from each other along the polar axis are independent of lens age, but their distance from the lens surface increases with increasing age. Upon accommodation, the distance between the two zones increases while all other distances along the polar axis remain unchanged, indicating that, as in the human, overall alterations in rhesus lenticular shape and thickness with accommodation primarily reflect changes in the shape of the central region. The curvature of each zone becomes sharper in a linear fashion with accommodation, with the slope of the relationship being similar to those for lens surfaces.

Accommodation, Ocular↗
PubMed1988

Electron microscopy of native and reconstituted alpha crystallin aggregates.

The size and shape of native alpha crystallin aggregates extracted at either 4 degrees C (alpha c-crystallin) or 37 degrees C (alpha m-crystallin), were compared with each other, as well as with aggregates reconstituted from either pure alpha A or alpha B subunits using electron microscopy. The alpha B aggregates were the most uniform in size (about 9 nm in diameter) and the best stained. alpha A particles were about the same size as the alpha B, but the population distribution was broader and some indication of interparticle association was observed. alpha c particles exhibited a bimodal distribution, with one peak greater than the reconstituted particles and the other about the same size; alpha m was smaller than the reconstituted structures.

Animals↗
PubMed1987

A possible structure for alpha-crystallin.

alpha-Crystallin, the major protein of the mammalian eye lens, is found in vivo as a multimeric aggregate composed of two closely related subunits whose molar ratio is widely variable from species to species. Attempts to determine the arrangement of the subunits within the aggregate, or even to determine the size of the aggregate and the number of subunits composing it, have not resulted in general agreement. Because of the variability in alpha-crystallin particle size, the apparent dependence of this parameter on certain environmental factors (e.g. temperature), the absence of a specific requirement for either alpha-crystallin isoform in aggregation, and the sharp division in the amino acid sequence between a strong hydrophobic region and a sharply hydrophilic one, it is suggested that the alpha-crystallin aggregate has the properties of a protein micelle. This hypothesis is consistent with what is known of the alpha-crystallin molecule and aggregate, and can be tested experimentally. If this hypothesis is shown to be true, then alpha-crystallin will be the first example of a naturally occurring protein micelle.

Animals↗
PubMed1987

Slit-lamp studies of the rhesus monkey eye. I. Survey of the anterior segment.

Slit-lamp photographic studies of 144 caged rhesus monkeys, aged 2 months to 35 years, show age-related changes in anterior-chamber depth, lens thickness, anterior and posterior curvatures of the lens, and location of the posterior lens surface relative to the anterior corneal surface. For these parameters, as well as for those measured by other techniques, a difference in slope magnitude and (or) slope sign was found between the growth phase which lasts for 5-6 years, and the adult phase (greater than 5-6 years). Age-related changes in the adult rhesus eye are qualitatively similar in almost all aspects to those observed in the human eye, indicating that the rhesus is a good animal model for the study of human loss of accommodative amplitude.

Aging↗
PubMed1987

Slit-lamp studies of the rhesus monkey eye: II. Changes in crystalline lens shape, thickness and position during accommodation and aging.

Changes in crystalline lens shape and axial thickness, anterior chamber depth and anterior cornea-posterior lens distance during accommodation induced by corneal iontophoresis of carbachol or electrical stimulation of the Edinger-Westphal nucleus were studied in 25 living, surgically aniridic rhesus monkey eyes, aged 1-25 years. Intraocular distances and anterior and posterior lens surface curvatures were evaluated from slit-lamp Scheimpflug photographs; distances were also determined by A-scan ultrasonography. With increasing accommodation, both lens surfaces become more sharply curved, the lens thickens and the anterior chamber shallows, while the posterior lens surface remains fixed relative to the cornea. Within statistical limits, the respective curvature and distance changes are the same for a given dioptric accommodation induced by either stimulation technique. The respective intraocular distance-accommodation relationships are identical whether derived from photographic or ultrasonographic measurements. Temporal and contralateral reproducibility of all measurements is excellent. Each parameter-accommodation relationship is strikingly linear in all eyes, although above 20 D the slopes of the lens surface curvature-accommodation relationships may have decreased. The curvature change per D of accommodation averages approximately 20% more for the posterior than for the anterior lens surface. There is relatively little interindividual variation in the slope of each relationship despite the significant interindividual differences in age and accommodative amplitude, indicating that the relationships are independent of age. However, when extrapolated back to the non-accommodated resting state, the data indicate that the lens thickens, both its surfaces become more sharply curved, and the anterior chamber shallows with age in adult greater than 5 years, while opposite trends are seen in younger animals.

Accommodation, Ocular↗
PubMed1984

Analysis of human crystalline lens curvature as a function of accommodative state and age.

Slit-lamp photographs from four human subjects, aged 11, 19, 29, and 45 were reanalyzed using computer-based digitization and curve-fitting methods in order to obtain more complete information on internal lens curvature changes during accommodation. All discernible curves (N = 742) could be fit to parabolas with chi 2 less than or equal to 0.001 irrespective of lens age, accommodative state, or curve location within the lens. For each lens, the coefficients of the parabolas, when displayed in graphic form, exhibit a linear relationship between location within the lens and the coefficient of the chi 2 term. The slope of this line remains unchanged over accommodation for a given lens, but is shifted in position. The slope changes as a function of age. The age 45 lens exhibits these characteristics to a limited extent only, the differences possibly related to the development of presbyopia. The further a given curve is located from the lens surface, the smaller the region of its arc that can be considered approximately circular. A roughly hourglass figure is generated by these circular bounds; the waist of the hourglass decreases with increasing accommodation, since changes in radius of curvature with accommodation are more pronounced internally. Calculations of arc lengths as a function of increasing accommodation indicate that these lengths change very little over the entire accommodative range.

Accommodation, Ocular↗
PubMed1983

A model for accommodation in the young human eye: the effects of lens elastic anisotropy on the mechanism.

An explanation of the mechanism of visual accommodation depends heavily on understanding the mechanical properties of the lens, as well as the way in which its shape is altered in small accommodative changes. An initial attempt to relate these properties to a mechanism has already been performed (Koretz and Handelman, 1982) for the young (age 11 yr) human lens, using certain simplifying assumptions (spherical curvature on the anterior lens surface and elastic isotropy). However, since it has been shown that the lens behaves as an anisotropic body, the previous treatment has been extended to include the variation of lens elastic properties in the polar and radial directions. With this modified representation, it is found that only one combination of elastic constants is consistent with the generally accepted qualitative theory of accommodation and with clinical data on the accommodative range of the emmetropic age 11 human eye. For this unique solution of the equations, however, the general mechanism already suggested by us, which includes support by the vitreous and alteration of the magnitude and angle of application of zonular force with accommodation, remains little changed.

Accommodation, Ocular↗
PubMed1982

Model of the accommodative mechanism in the human eye.

The crystalline lens of the age 11 human eye has been modelled mathematically, using simplified assumptions about lens curvature, internal organization and elasticity. From this representation, expressions for description of strain and stress during accommodation have been obtained. Solution of these equations indicates that the lens capsule acts as a force distributor, spreading tension applied by the suspensory apparatus evenly over the surface of the underlying lens material. It also becomes clear that the vitreous body provides an essential support function during the accommodative process. Finally, the relative contribution of lens-associated structures has been determined for five different values of the Poisson ratio. In order for accommodation to occur by relaxation of zonular tension, this value must be greater than 0.38; with an additional constraint of the net axial force equalling zero during a small accommodative change, the Poisson ratio equals 0.46.

Accommodation, Ocular↗
PubMed1982

The aggregation characteristics of column-purified rabbit skeletal myosin in the presence and absence of C-protein at pH 7.0.

The aggregation properties of column-purified rabbit skeletal myosin at pH 7.0 were investigated as functions of ionic strength, protein concentration, and time. Filaments prepared by dialysis exhibited the same average length and population distribution at 0.10 and 0.15 M KCl at protein concentrations greater than 0.10 mg/ml; similar results were obtained at .0.20 M KCl, although average filament length was approximately 0.5 micrometer shorter. Once formed, these length distributions remained virtually unchanged over an 8-d period. At and below 0.10 mg/ml, average filament length decreased as a function of protein concentration; filaments prepared from an initial concentration of 0.02 mg/ml were half the length of those prepared at 0.2 mg/ml. Filaments prepared by dilution exhibited a sharp increase in average length as the time-course increased up to 40 s, then altered only slightly over a further period of 4 min. Addition of C-protein in a molar ratio of 1-3.3 myosin molecules affected most of these results. Average filament length was affected neither by ionic strength nor by initial protein concentration down to 0.04 mg/ml or over an 8-d period. Filaments formed by dilution in the presence of C-protein exhibited a constant average length and hypersharp length distribution over variable time courses up to 7 min. It is possible that C-protein acts to stabilize the antiparallel intermediate during filamentogenesis, and may also affect subunit addition to this nucleus.

Animals↗
PubMed1982

Structural studies of isolated native thick filaments from rabbit psoas muscle with AMP deaminase decoration.

AMP deaminase (adenylate deaminase; AMP aminohydrolase, EC 3.5.4.6), a large flat tetrameric enzyme found in skeletal muscle, binds strongly and specifically to the subfragment-2 region of rabbit skeletal muscle myosin. This allows its use as a structural probe in myosin and myosin rod aggregation studies. When mixed with a preparation of isolated native thick filaments, AMP deaminase decorates the entire filament backbone except for the central bare zone. Binding is particularly ordered in the banded region, where 11 stripes of about 43-nm spacing on either side of the bare zone have been observed in studies of isolated A-bands. No systematic absence of deaminase is seen here, indicating that the presence of the C-protein and H-protein bands does not make the binding site inaccessible to the tetramer. Optical diffraction patterns of the decorated filaments show the expected 42.9-nm periodicities and a reflection indexing at 28.6 nm. Because of the bulkiness of the tetramer relative to the number of available binding sites at each 14.3-nm interval along the filament shaft, the helix net is being sampled at a lower frequency than is the underlying myosin organization. As a result, reflections on layer lines other than orders of 42.9 nm are also observed (e.g., 57.2); these reflections strongly indicate a structure based on a 12/1 primitive helix. The results appear to eliminate the symmetric double two-fold and three-fold models of thick filament structure but are consistent with an asymmetric four-fold structure.

AMP Deaminase↗
PubMed1980

Adenylate deaminase binding to synthetic thick filaments of myosin.

Adenylate deaminase (AMP deaminase; AMP aminohydrolase, EC 3.5.4.6), a tetrameric enzyme found at particularly high concentrations in skeletal muscle, has previously been shown to bind strongly to the subfragment-2-portion of myosin in vitro and to the ends of the A band in vivo. It is shown here that when adenylate deaminase is dialyzed with skeletal myosin during formation of synthetic filaments at pH 7.0 it decorates the filament at 14.3-nm intervals, presumably in the region of exposed backbone between crossbridge levels. Optical diffraction of the aggregates reveals both enhancement of reflections arising from underlying myosin organization and other reflections arising from adenylate deaminase arrangement on the filament surface. Adenylate deaminase can thus be used as a specific label in the study of myosin presence and organization.

AMP Deaminase↗