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71.
Purification and crystallization of complexes modeling the active state of the fragile histidine triad protein 总被引:1,自引:0,他引:1
Brenner C; Pace HC; Garrison PN; Robinson AK; Rosler A; Liu XH; Blackburn GM; Croce CM; Huebner K; Barnes LD 《Protein engineering, design & selection : PEDS》1997,10(12):1461-1463
Fragile histidine triad protein (Fhit) is a diadenosine triphosphate
(ApppA) hydrolase encoded at the human chromosome 3 fragile site which is
frequently disrupted in tumors. Reintroduction of FHIT coding sequences to
cancer cell lines with FHIT deletions suppressed the ability of these cell
lines to form tumors in nude mice even when the reintroduced FHIT gene had
been mutated to allow ApppA binding but not hydrolysis. Because this
suggested that the tumor suppressor activity of Fhit protein depends on
substrate-dependent signaling rather than ApppA catabolism, we prepared two
crystalline forms of Fhit protein that are expected to model its
biologically active, substrate-bound state. Wild-type and the His96Asn
forms of Fhit were overexpressed in Escherichia coli, purified to
homogeneity and crystallized in the presence and absence of ApppA and an
ApppA analog. Single crystals obtained by vapor diffusion against ammonium
sulfate diffracted X-rays to beyond 2.75 A resolution. High quality native
synchrotron X-ray data were collected for an orthorhombic and a hexagonal
crystal form.
相似文献
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TM Ortega EC Feldman RW Nelson N Willits LD Cowgill 《Canadian Metallurgical Quarterly》1996,209(10):1724-1729
OBJECTIVE: To determine prevalence and severity of systemic arterial hypertension and proteinuria in dogs with naturally developing hyperadrenocorticism and to determine whether these abnormalities resolve with adequate management of the disease. DESIGN: Case series and cohort study. ANIMALS: 77 dogs with naturally developing hyper-adrenocorticism examined once; 15 dogs examined before and after treatment. RESULTS: Among dogs examined only once, hypertension was diagnosed in 21 of 26 dogs with untreated pituitary-dependent hyperadrenocorticism (PDH), 17 of 21 with inadequately controlled PDH, 8 of 16 with well-controlled PDH, 10 of 10 with an untreated adrenocortical tumor, and 0 of 4 that had undergone adrenalectomy because of an adrenocortical tumor. Untreated dogs and dogs with inadequately controlled PDH had significantly higher blood pressures than did other dogs. Proteinuria was documented in 12 of 26 dogs with untreated PDH, 5 of 16 with inadequately controlled PDH, 3 of 14 with well-controlled PDH, 5 of 8 with an untreated adrenocortical tumor, and 1 of 3 that had undergone adrenalectomy. Dogs with untreated PDH and dogs with an untreated adrenocortical tumor had higher urine protein/creatinine ratios than did dogs with well-controlled PDH. Among dogs evaluated before and after treatment, blood pressure and urine protein/creatinine ratio did not change in 8 dogs with inadequately controlled hyperadrenocorticism, but decreased in 7 dogs with well-controlled disease. CLINICAL IMPLICATIONS: Results suggest that systemic hypertension and proteinuria are common in dogs with untreated hyperadrenocorticism and that successful treatment of hyperadrenocorticism will result in resolution of these abnormalities in many, but not all, dogs. 相似文献
75.
LD Boada L Fernández M Zumbado OP Luzardo R Chirino BN Díaz-Chico 《Canadian Metallurgical Quarterly》1996,279(3):1123-1129
Male rat liver microsomes contain a [3H]dexamethasone binding site, capable of binding glucocorticoids and progesterone. We have shown previously that the 17 alpha-alkylated androgen, stanozolol, can inhibit the [3H]dexamethasone binding to microsomes through a negative allosteric mechanism, which gives rise to the possibility of its interaction with a different binding site. In this study, the existence of a single-saturating binding site, capable of binding the radioactive steroid with a maximum number of the specific binding site of 49 +/- 2 pmol/mg of protein and a Kd of 37 +/- 1.3 nM was demonstrated by using [3H]stanozolol. In competition experiments, only stanozolol and danazol were able to compete with [3H]stanozolol for its binding to microsomes, among more than 60 steroids and other compounds tested. The binding of [3H]stanozolol was depressed after protease treatment of the microsomes, or after the administration of cycloheximide to adult male rats for 24 hr, which suggest its proteic nature. The [3H]stanozolol binding site was detected in many tissues of the rat, with the highest concentrations being found in the liver. It was detected from birth, increasing afterward in concentration and reaching a peak at 2 to 3 months of age. This is the first experimental verification of the existence in liver microsomes of a specific binding site for some 17 alpha-alkylate androgens, such as stanozolol and danazol, different from the androgen receptor or the [3H]dexamethasone binding site. 相似文献
76.
FB Mochales RB Castelló JC Marqués LD Rodríguez XG Alemany JA Rodríguez EP Beato BP Gorricho EP Trallero GP Pastor 《Canadian Metallurgical Quarterly》1995,69(6):445-461
This document represents the recommendations of a panel of Spanish experts on antibiotic use and resistance. In a Task Force, under the auspices of the Spanish Ministry of Health and Consumer Affairs that took place in 1994 in Madrid, the members were gravely concerned about the national increase in antibiotic resistance. They analysed the development, evolution and spread of antibiotic resistance among community-acquired human bacterial pathogens in Spain, its relation with antibiotic consumption, and they proposed future surveillance strategies for monitoring the patterns of antibiotic use and consumption. Success will require a collective action among the producers (pharmaceutical industry), prescribers (doctors, veterinarians), dispensers (pharmacists), and consumers (patients). Two similar documents have been recently published by the American Society of Microbiology and the World Health Organization showing the global concern about this topic. 相似文献
77.
S Abdi LD Traber DN Herndon CS Rogers DL Traber 《Canadian Metallurgical Quarterly》1995,293(4):475-481
We studied the effects of ibuprofen on bronchial blood flow and myocardial function after inhalation injury. Sheep (n = 12) were chronically instrumented with cardiovascular and pulmonary catheters. After 5 days of recovery period, baseline data were collected and the sheep were divided into two groups. Group S (n = 6) were insufflated with 48 breaths of cotton smoke; while group I (n = 6) were pretreated with ibuprofen (12mg/kg bolus followed by 3 mg/kg/h continuous infusion for 24 h) and challenged with the same dose of smoke. All the animals were studied for 24h. Bronchial blood flow increased significantly in both groups throughout the experimental period; while stroke volume as well as right and left ventricular stroke work indices of both groups were significantly decreased (group I worse than group S) in the second half of the experimental period. These data suggest that vasodilatory prostaglandins do not play a major role in the bronchial vascular response to smoke inhalation injury and myocardial depression seen post injury is worse in animals treated with ibuprofen. 相似文献
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H Kino H Inaba N Van De L Van Chau DT Son HT Hao ND Toan LD Cong M Sano 《Canadian Metallurgical Quarterly》1998,29(2):250-254
We present the spatial structure of binase, a small extracellular ribonuclease, derived from 1H-NMR* data in aqueous solution. The total of 20 structures were obtained via torsion angle dynamics using DYANA program with experimental NOE and hydrogen bond distance constraints and phi and chi1 dihedral angle constraints. The final structures were energy minimised with ECEPP/2 potential in FANTOM program. Binase consists of three alpha-helices in N-terminal part (residues 6-16, 26-32 and 41-44), five-stranded antiparallel beta-sheet in C-terminal part (residues 51-55, 70-75, 86-90, 94-99 and 104-108) and two-stranded parallel beta-sheet (residues 22-24 and 49-51). Three loops (residues 36-39, 56-67 and 76-83), which play significant role in biological functioning of binase, are flexible in solution. The differences between binase and barnase spatial structures in solution explain the differences in thermostability of binase, barnase and their hybrids. 相似文献
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