PubMed Health⌕ Search

Biomedical subjects

I Pilz

Publications and source records attributed to I Pilz.

At least 19 recordsLinked to original sources
PubMed1991

Deletion of the linker connecting the catalytic and cellulose-binding domains of endoglucanase A (CenA) of Cellulomonas fimi alters its conformation and catalytic activity.

The Pro-Thr box is a linker of 23 amino acids ((PT)4T(PT)7) connecting the catalytic domain and the cellulose-binding domain (CBD) of endoglucanase A (CenA) from the bacterium Cellulomonas fimi. Deletion of the Pro-Thr box alters the conformation of CenA by changing the relative orientation of the catalytic domain and the CBD. The tertiary structures of the catalytic domain and the CBD appear to be unchanged. The change in conformation reduces the catalytic efficiency of the enzyme and masks one of two protease-sensitive sites between the domains. The deletion does not affect the adsorption of the enzyme to microcrystalline cellulose, but it does affect its desorption from cellulose. The results suggest that the Pro-Thr box in CenA has an extended, kinked, and rigid conformation.

Actinomycetales↗
PubMed1991

Small-angle X-ray study on the quaternary structure of erythrocruorin from Caenestheria inopinata.

The erythrocruorin of the clam shrimp Caenestheria inopinata was studied in sodium phosphate buffer at pH 6.8 by small-angle X-ray scattering. The following molecular parameters were determined: radius of gyration 4.77 +/- 0.05 nm, maximum dimension 14.0 +/- 0.5 nm and a volume of 640 +/- 40 nm3. A model which fits the experimental data well is presented. The model is composed of 10 subunits arranged symmetrically in two layers, whereby five subunits are always forming a ring.

Animals↗
PubMed1990

The tertiary structure of a bacterial cellulase determined by small-angle X-ray-scattering analysis.

CenA from Cellulomonas fimi is a beta-1,4-endoglucanase that binds tightly to cellulose. X-ray-scattering analyses show that the enzyme is tadpole-shaped: the previously identified catalytic and cellulose-binding domains comprise the head and tail respectively. It appears that this structural and functional organization is common to several cellulases from bacteria and fungi.

Actinomycetales↗
PubMed1990

Quaternary structure of the giant extracellular hemoglobin of the leech Macrobdella decora.

The molecular dimensions of the extracellular hemoglobin of the leech Macrobdella decora, determined by scanning transmission electron microscopy were 29.8 nm x 19.5 nm (diameter x height) for negatively stained specimens. Measurements of molecular mass (Mm) of unstained specimens with the microscope gave Mm = 3560 +/- 160 kDa. Small-angle X-ray scattering measurements gave a diameter of 28.0(+/- 0.5) nm, radius of gyration 10.5(+/- 0.2) nm and volume 7500(+/- 300) nm3. The hemoglobin had no carbohydrate and its iron content was found to be 0.23(+/- 0.02)% (w/w), corresponding to a minimum Mm of 24,000(+/- 1300) kDa. SDS/polyacrylamide gel electrophoresis of the unreduced hemoglobin showed that it consisted of three subunits, which have apparent Mm values of 12 (1), 25 (2) and 29 kDa (3). The reduced hemoglobin consisted of four subunits, I (12 kDa), II (14 kDa), III (26 kDa) and IV (30 kDa). Subunit 1 corresponded to subunit I, subunit 2 to subunits III and IV and subunit 3 to subunit II. Partial N-terminal sequences were obtained for subunit 1, the two chains of subunit 2 and one of the two chains of subunit 3, suggesting that the hemoglobin consists of at least five different polypeptide chains. The percentage fraction of the three unreduced subunits was determined by densitometry of SDS/polyacrylamide gel patterns and quantitative determination of Coomassie R-250 dye bound to the individual bands in reduced and unreduced patterns to be, monomer (subunit I) : non-reducible subunit (subunit 2) : reducible dimer (subunit 3) = 0.35 : 0.29 : 0.35 (S.D. = +/- 0.05). This corresponded to a stoichiometry of 74 +/- 11 : 37 +/- 5 : 38 +/- 6, assuming the molecular masses to be 17 kDa, 30 kDa and 34 kDa, taking into account the anomalously high mobility of annelid globins in SDS-containing gels. The stoichiometry calculated from the amino acid compositions of the hemoglobin and the three subunits was 82 +/- 12 : 29 +/- 4 : 40 +/- 8. Gel filtration of the hemoglobin at pH 9.8, at neutral pH subsequent to dissociation at pH 4 and at neutral pH in the presence of urea and Gu.HCl provided no evidence for the existence of a putative 1/12 of the whole molecule (Mm approx. 300 kDa). Furthermore, the largest subunits obtained had Mm of 60 to 100 kDa and had a much decreased content of subunit 2, suggesting that the hemoglobin was not a simple multimeric protein. Three-dimensional reconstruction from microscope images provided a model of Macrobdella hemoglobin that is very similar to the reconstruction of Lumbricus hemoglobin: the radial mass distribution curves are virtually superimposable.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acid Sequence↗
PubMed1989

Structural changes in cellobiohydrolase I upon binding of a macromolecular ligand as evident by SAXS investigations.

Xylan from Rhodymenia palmata binds to the cellobiohydrolase I from Trichoderma reesei (CBH I) or its core protein, inhibiting their activity. Adsorption onto microcrystalline cellulose (Avicel) is reduced approximately 30% for intact CBH I and nearly 50% for the core, whereas the effects with cellobiose are negligible. Structural changes concomitant with this binding are studied in solution by small angle X-ray scattering. In the "tadpole" structure typical for the CBH I [Abuja et al., 1988] the lengthening of the tail part is the most salient observation when xylan is present which accounts for an increase in Dmax (18.0 to 22.0 nm) and radius of gyration (4.74 to 5.18 nm). When xylan binds to the core the radius of gyration remains nearly unchanged. Here a model can be constructed showing a xylan molecule on the surface of the core protein near the tail part.

Adsorption↗
PubMed1988

Domain structure of cellobiohydrolase II as studied by small angle X-ray scattering: close resemblance to cellobiohydrolase I.

Evidence for a domain structure of cellobiohydrolase II (CBH II, 58 kDa) from Trichoderma reesei (Teeri et al., 1987; Tomme et al., 1988) is corroborated by results from SAXS experiments. They indicate a 'tadpole' structure for the intact CBH II in solution (Dmax = 21.5 +/- 0.5 nm; Rg = 5.4 +/- 0.1 nm) and a more isotropic, ellipsoid shape for the core protein (Dmax = 6.0 +/- 0.3 nm; Rg = 2.1 +/- 0.1 nm). The latter was obtained by partial proteolysis with papain which cleaves the native CBH II to give two fragments (Tomme et al., 1988): the core (45 kDa) with the active (hydrolytic) domain and a smaller fragment (11 kDa) coinciding with the tail part of the model and containing the binding domain for unsoluble cellulose. This peptide fragment is conserved in most cellulolytic enzymes from Trichoderma reesei (Teeri et al., 1987). It contains a conserved region (block A) and glycosylated parts (blocks B and B' duplicated and located N-terminally in CBH II). In spite of different domain arrangements in CBH I (blocks B-A at C-terminals) SAXS measurements (Abuja et al., 1988) indicate similar tertiary structures for both cellobiohydrolases although discrete differences in the tail parts exist.

Binding Sites↗
PubMed1988

Small angle X-ray scattering study on bovine porphobilinogen synthase (5-aminolaevulinate dehydratase).

The quaternary structure of the native (zinc) porphobilinogen synthase (5-amino-laevulinate dehydratase) from bovine liver and its lead-substituted derivative is studied in solution by small angle X-ray scattering. In spite of the profound inhibitory effect of lead ions in the enzyme they do not produce a change in the quaternary structure detectable by small angle X-ray scattering. The most important molecular parameters of the native enzyme were found to be: radius of gyration Rg = 4.04 +/- 0.04 nm and maximum dimension Dmax = 12.0 +/- 0.5 nm. The corresponding values for the lead derivative are: Rg = 4.26 +/- 0.1 nm and Dmax = 12.5 +/- 0.5 nm. The quaternary structure of the enzyme in solution is described by a model, which fits the experimental scattering and distance distribution function.

Animals↗
PubMed1987

Epitope localization in antigen-monoclonal-antibody complexes by small-angle X-ray scattering. An approach to domain organization in the beta 2 subunit of Escherichia coli tryptophan synthase.

Each polypeptide chain of the beta 2 subunit of Escherichia coli tryptophan synthase (EC 4.2.1.20) is made of two domains, F1 (N-terminal) and F2 (C-terminal). To determine the relative position of these domains in the native protein, complexes between beta 2 and Fab fragments from two monoclonal antibodies, one specific for F1 (68-1) and the other for F2 (93-6), have been prepared and purified. Small-angle X-ray scattering measurements have been made on solutions of each complex. From the experimental scattering curves obtained, computer modeling leads to structural models of the two beta 2-Fab complexes. Though relatively low, the resolution of these models allows the localization on beta 2 of the antigenic sites recognized by the two antibodies, to show that the C-terminal F2 domains lie at the distal ends of the elongated beta 2 protein, and to show how steric hindrance prevents beta 2, though structurally and functionally dimeric, from binding more than one Fab 93-6 fragment per dimer.

Antigen-Antibody Complex↗
PubMed1986

[Hyalinosis cutis et mucosae--Urbach-Whiete syndrome. Review and case report].

We present the Urbach-Whiete-Syndrome as a rare cause of hoarseness in children. As such patients often present initially to the E.N.T. surgeon, we should like to draw attention to this autosomal recessive condition. Only symptomatic treatment is possible, and genetic counselling should be considered. The diagnosis of the disease should be as prompt as possible.

Abnormalities, Multiple↗
PubMed1979

[Rickets prophylaxis by daily administration of vitamin D tablets (author's transl)].

The efficiency of a daily vitamin D prophylaxis with tablets ("Laevovit D3"), containing 1000 I.U. cholecalciferol was studied in newborn babies (most of them "prematures") over a period of about 5 weeks. The blood-values of 25-hydroxycholecalciferol were taken as parameter and determined 1 week and 5 weeks after the start of vitamin D prophylaxis. The data obtained were--with very few exceptions-- in a range considered to be protective against development of vitamin D deficiency. No significant difference could be found comparing vitamin D prophylaxis with these "microtablets" vs. liquid vitamin D preparations.

Female↗
PubMed1978

[Plasma 25-hydroxycholecalciferol after daily vitamin D administration in comparison with massive single-dose prophylaxis (author's transl)].

The effect of daily administration of 1200 IU vitamin D3 was compared with the response to a single oral dose of 200,000 IU in 21 young infants by determination of the plasma 25-OHCC levels. After about one week the mean value recorded in group 1 was 27 +/- 13 ng/ml; the corresponding value in group 2 was 127 +/- 78.4 ng/ml. At the second control examination about one month subsequently, the mean values were 61 +/- 26.2 ng/ml and 104 +/- 69.0 ng/ml respectively. The wide range of values found in the single-dose group clearly demonstrates the advantage of daily vitamin D administration for routine prophylaxis against rickets in infancy.

Administration, Oral↗
PubMed1978

Small-angle X-ray studies of a human immunoglobulin M.

The conformation of a Waldenström immunoglobulin M (IgM) with antibody-like activity for X-ray contrast media, based on 3-amino-2,4,6-triiodobenzoic acid, was studied by small-angle X-ray scattering. The radius of gyration was determined as 12.1 nm, the maximum distance 35 nm, the volume 1900 nm3. A flat star-shaped model was found to be equivalent in scattering. Aggregation of IgM molecules seems to take place as side-by-side combinations of single molecules, manifesting itself as a relatively large increase of the radius of gyration and unchanged thickness of the flat aggregates.

Humans↗
PubMed1977

[Small-angle X-ray-scattering investigation and structural-model study of the fatty-acid synthetase from pig liver (author's transl)].

The structure of the fatty acid synthetase from pig liver was studied on models based upon structural and functional properties selected from pertinent results available from numerous investigations carried out with fatty acid synthetases from this and other sources. When comparing small-angle X-ray-scattering curves calculated with these models and curves obtained from small-angle X-ray-scattering experiments carried out with the pig-liver enzyme, we tried to select a model which would lead to an acceptable correlation between the calculated and the experimental curves and at the same time fulfil the known structural and functional requirements. The comparison of the curves was started with a model of low complexity. The observed discrepancy, together with arguments from the structural and the functional properties, helped decide which is the next most reasonable model to be considered. This procedure was repeated for five models of increasing complexity. In the model which led to the best fit the multienzyme complex is composed of two halves in an assymetric conformation including hollow spaces. This highly anisotropic model would imply that the two halves change their conformation each time a synthetic cycle is completed and that the growing fatty acid is handed over from one half to the other.

Animals↗
PubMed1977

Studies by small-angle X-ray scattering of the quaternary structure of the beta-haemocyanin of Helix pomatia.

Helix pomatia beta-haemocyanin was studied in solution by small-angle X-ray scattering. The following molecular parameters were determined: molecular weight = 9.02 X 10(6), volume = 14000 nm3, radius of gyration = 18.4 nm, radius of the spherical subunits = 2.5 +/- 0.2 nm. With these data, and with information of dissociation products described in a former paper, a model of the molecule was built whose theoretical scattering curve showed good agreement with the experimental one. The model consists of 160 spherical subunits of a radius of 2.5 nm; 12 rings each built up of 10 spheres form the outer wall of a hollow cylinder; 20 subunits are situated at the inner side of each end.

Animals↗
PubMed1977

On the conformation of serine-specific transfer RNA. Studies by small-angle X-ray scattering and ultraviolet absorption of the molecule in solution.

The scattered X-ray intensities from dilute solutions of tRNASer (yeast) in 0.1 M Soerensen buffer at pH 7.0 were measured at 25 degrees C. The radius of gyration, molecular weight and volume were determined. A model equivalent in scattering is given. The change of the conformation of tRNASer by heating was followed by small-angle X-ray measurements and ultraviolet absorption in a temperature range 20-70 degrees C. The molecule begins to unfold at about 40 degrees C and 70 degrees C has a random coil conformation. Addition of magnesium stabilizes the tRNASer molecule. The reversibility of the melting process was also studied by both methods. An interesting effect was found by ultraviolet absorption: by heating the tRNASer solutions to 55 degrees C and 60 degrees C and subsequently slowly cooling, the melting curves lie at higher absorption values than the corresponding cooling curves. The small-angle data and optical properties of tRNASer are compared with those of tRNAPhe which has already been thoroughly investigated.

Binding Sites↗