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1.
Epitope analysis and molecular modeling reveal the topography of the C-terminal peptide of the beta-subunit of human chorionic gonadotropin. 总被引:2,自引:0,他引:2
N Venkatesh S Krishnaswamy S Meuris G S Murthy 《European journal of biochemistry》1999,265(3):1061-1066
Human chorionic gonadotropin (hCG) belongs to a family of heterodimeric glycoprotein hormones that share a common alpha-subunit and a hormone-specific beta-subunit. Among the gonadotropin beta-subunits, greater than 85% homology exists between lutropin (hLH)beta and hCGbeta in their first 114 amino acid residues. However, unlike hLHbeta, hCGbeta contains a 31-amino acid hydrophilic stretch at its carboxyl end (CTPbeta: C-terminal peptide). Although the crystal structure of deglycosylated hCG has been solved, the topography of CTPbeta remains unknown. In this study, we have attempted to define the topology of CTPbeta using mAb probes. We investigated three epitopes on hCGalpha, which are hidden in the hCGalphabeta dimer. However, these epitopes are not hidden in hLH, which has a similar subunit interface to that of hCG, but lacks CTPbeta. This suggested that these epitopes are not masked at the subunit interface of hLH or hCG. Hence, we hypothesized that, in the case of hCG, these epitopes are masked by the CTPbeta. Consistent with this view, several treatments of hCG that removed CTPbeta unmasked these epitopes and enhanced their reactivity with the corresponding mAbs. In order to localise the position of CTPbeta on the alpha-subunit, we used an epitope-mapping strategy [N. Venkatesh & G. S. Murthy (1997) J. Immunol. Methods 202, 173-182] based on differential susceptibility of epitopes to covalent modifications. This enabled us to predict the possible topography of CTPbeta. Further, we were also able to build a model of CTPbeta, completely independently of the epitope-mapping studies, using a homology-based modeling approach [S. Krishnaswamy, I. Lakshminarayanan & S. Bhattacharya (1995) Protein Sci. 4 (Suppl. 2), 86-97]. Results obtained from these two different approaches (epitope analysis and homology modeling) agree with each other and indicate that portions of CTPbeta are in contact with hCGalpha in the native hCG dimer. 相似文献
2.
OBJECTIVE: To determine the vaccination rate among infants discharged from a neonatal intensive care unit (NICU) and factors affecting that rate. DESIGN: Cross-sectional survey conducted when the children were 12 to 18 months of age. SETTING: NICU at the Royal University Hospital, Saskatoon, Sask. PARTICIPANTS: All 395 infants discharged from the NICU between Jan. 1 and June 30, 1992. MAIN OUTCOME MEASURES: Vaccination rate, ethnic background (native or non-native), place of residence (urban or rural), health status (number of days spent in the NICU), reasons for delay in or incomplete vaccinations (those involving parents'' responsibility, infant illness or contraindications). RESULTS: Of the 395 infants, 20 (5.0%) had died and incomplete information was available for 30 (7.6%). Complete data were available for 345 (87.3%). Of the infants for whom data were available, 8 (2.3%) had never been vaccinated and 142 (41.2%) had a delayed vaccination schedule or had not completed their scheduled vaccinations. Only 195 (56.6%) of the infants had received a full vaccination series. Non-native ethnic background was a predictor of completed vaccinations (odds ratio [OR] 5.40, 95% confidence interval [CI] 3.05 to 9.52). In a univariate model, urban area of residence was not a significant predictor of vaccination status, but when ethnic background was controlled for in a multivariate logistic regression analysis, urban area of residence was found to be inversely associated with completed vaccinations (OR 0.34, 95% CI 0.15 to 0.79). The number of days the child had spent in the NICU was not a significant predictor of vaccination status. CONCLUSION: The vaccination rate of infants discharged from the NICU is not optimal. Urban native children appears to be at risk of not being vaccinated. Non-native infants are five times more likely than native infants to have completed all of their scheduled vaccinations. Methods to improve the rate of completed vaccinations, especially for native children, must be sought and tested. 相似文献
3.
Membrane penetration of bovine factor V and Va detected by labeling with 5-iodonaphthalene-1-azide 总被引:2,自引:0,他引:2
M F Lecompte S Krishnaswamy K G Mann M E Nesheim C Gitler 《The Journal of biological chemistry》1987,262(5):1935-1937
The membrane-binding properties of Factor V and Factor Va were investigated using the lipophyllic, photoactivable probe 5-[125I]iodonaphthalene-1-azide. In the presence of vesicles composed of 75% phosphatidylcholine and 25% phosphatidylserine, both Factor V and Va were found to be labeled by the probe. The label was almost exclusively localized to the carboxyl-terminal-derived component E of Factor Va. The results are consistent with the interpretation that component E is the membrane binding subunit of Factor Va and that the interaction between Factor V or Factor Va and the membrane involves the penetration of the protein into the lipid bilayer. 相似文献
4.
Ashwini Deshpande K. Sankaran S. F. D'Souza G. B. Nadkarni 《Biotechnology Techniques》1987,1(1):55-58
Summary A relatively simple method has been described for the rapid purification of D-amino acid oxidase from Trigonopsis variabilis by hydrophobic chromatography on Phenyl-Sepharose CL-4B and negative adsorption on DEAE-cellulose. The purified enzyme had a specific activity of 22–24 units at 25°C and exhibited three bands on enzymatic staining. 相似文献
5.
To determine whether a diurnal rhythm exists in neonates admitted to neonatal intensive care units where there is continuous artificial lighting and periodic nursing and medical care, plasma cortisol, adrenocorticotropin (ACTH), and beta-endorphin concentrations were measured in two groups of infants and in adult human volunteers. As expected, a diurnal rhythm was seen in adults. A diurnal rhythm was also found for cortisol and endorphin levels in neonates (3 to 4 days postnatally) with minimal stress and in infants who were clinically severely stressed. There was not a significant difference between the morning and afternoon concentrations of ACTH in these infants, but the afternoon concentrations were lower than the morning''s, as would be expected. We found that a diurnal rhythm does exist in neonates within the first few days of postnatal life and that the continuous lighting and medical and nursing interventions do not interfere with this rhythm. 相似文献
6.
The most abundant anhydrase isoenzyme from the erythrocyte of Indian buffalo has been purified using affinity gel and DEAE-cellulose ion-exchange columns and single crystals suitable for X-ray diffraction studies have been obtained. The unit cell dimensions are a = 46.8 A, b = 104.5 A, c = 60.4 A, beta = 91.2 degrees and the space group is P2(1), with two molecules per asymmetric unit. 相似文献
7.
Epidermal growth factor (EGF) enhances the induction of alpha-lactalbumin in mammary explants from pregnant and virgin rats in the presence of insulin (I), hydrocortisone (F) and prolactin (P). EGF also enhances the prolactin-independent induction of alpha-lactalbumin in tissue from pregnant rats and evokes prolactin-independent induction of alpha-lactalbumin in mammary tissue from virgin rats in the presence of I and F. Casein synthesis and galactosyltransferase activity are unaffected by EGF in the IFP-system, and are not induced in the IF-EGF-system. Multiplication stimulating activity, nerve growth factor, fibroblast growth factor and platelet-derived growth factor do not mimic the selective effects of EGF on rat alpha-lactalbumin. These influences of EGF on the differentiation of isolated rat mammary tissue are compared with those on mouse and rabbit tissue studied previously. 相似文献
8.
Mammary explants from pregnant rats can be induced in regard to casein synthesis and alpha-lactalbumin activity when cultured in the presence of hydrocortisone, prolactin and levels of insulin approaching physiological concentrations. No detectable induction occurs in the absence of insulin. Although epidermal growth factor and multiplication stimulating activity, in the presence of hydrocortisone, can maintain the initial level of NADH-cytochrome c reductase as well as insulin, neither can substitute effectively for insulin in the induction of the milk proteins. Proinsulin, nerve growth factor, platelet-derived growth factor and fibroblast growth factor are also ineffective substitutes for insulin in this regard. Whereas prolonged tissue exposure to multiplication stimulating activity, hydrocortisone and prolactin does not result in induction of alpha-lactalbumin activity, subsequent addition of insulin leads to prompt response. The results suggest that the ability of insulin to function as a unique, essential factor in the induction of rat milk proteins is independent of its cell-maintenance activity. Thus, in addition to its well established functions in metabolic processes, insulin appears to play a vital role in certain developmental processes. 相似文献
9.
Rabbit intestinal glucoamylase-maltase was examined in detail with respect to its molecular weight, sedimentation, diffusion and viscosity. It is a large asymmetrical molecule, with a molecular weight of 750 000-760 000. Its appearance under the electron microscope supports the idea that it is a long string (62.0 nm) consisting of eight beads of diameter 6.0 nm each and a surface-to-surface interbead distance of approx. 2.0 nm. The shape of the enzyme derived from its hydrodynamic behaviour by using the string-of-spherical-beads model originally proposed by Kuhn [(1932) Z. Phys. Chem. Abt. A 161, 1-32] and later modified by Shulman [(1953) J. Am. Chem. Soc. 75, 5846-5852] fits moderately well with the electron-microscopic picture. The beads might represent about six subunits, and the absence of sulphur from the enzyme and the inability to dissociate the enzyme by conventional methods indicate the possibility of unusual covalent cross-linking between the subunits and between the beads. 相似文献
10.
Résumé L'AcridienGesonula punctifrons
St?l attaque la Jacinthe d'eau (Eichhornia crassipes) dans plusieurs parties de l'Inde et est adapté à un habitat aquatique ou semi-aquatique. Les œufs, pondus dans les pétioles
des feuilles, éclosent en 3 à 4 semaines. Les larves se développent en 4 à 5 semaines et demi, avec 5 mues. Les adultes vivent
au maximum 4 à 5 mois en s'alimentant seulement de Jacinthe d'eau. En essais de laboratoire cet Acridien se nourrit activement
deCanna orientalis dont les tiges sont percées de trous pour la ponte par les femelles, mais aucun œuf n'est déposé. Quarante trois autres plantes
d'intérêt économique ont été expérimentées: plusieurs d'entre elles ont été l'objet d'attaques légères à modérées. L'analyse
de la littérature concernant les plantes-h?tes du genreGesonula et la distribution deG. punctifrons montrent que cette espèce s'est adaptée secondairement àE. crassipes. Bien que cet Acridien s'observe en grand nombre dans quelques régions, l'ensemble de ses dégats à la mauvaise herbe n'est
pas suffisant pour assurer le contr?le de celle-ci.
This research has been financed in part by a grant made by the United States Department of Agriculture under P. L. 480. 相似文献
This research has been financed in part by a grant made by the United States Department of Agriculture under P. L. 480. 相似文献