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11.
Summary The perdeuteration of aliphatic sites in large proteins has been shown to greatly facilitate the process of sequential backbone and side-chain 13C assignments and has also been utilized in obtaining long-range NOE distance restraints for structure calculations. To obtain the maximum information from a 4D 15N/15N-separated NOESY, as many main-chain and side-chain 1HN/15N resonances as possible must be assigned. Traditionally, only backbone amide 1HN/15N resonances are assigned by correlation experiments, whereas slowly exchanging side-chain amide, amino, and guanidino protons are assigned by NOEs to side-chain aliphatic protons. In a perdeuterated protein, however, there is a minimal number of such protons. We have therefore developed several gradient-enhanced and sensitivity-enhanced pulse sequences, containing water-flipback pulses, to provide through-bond correlations of the aliphatic side-chain 1HN/15N resonances to side-chain 13C resonances with high sensitivity: NH2-filtered 2D 1H-15N HSQC (H2N-HSQC), 3D H2N(CO)C/ and 3D H2N(COC/)C/ for glutamine and asparagine side-chain amide groups; 2D refocused H(N/)C/ and H(N/C/)C/ for arginine side-chain amino groups and non-refocused versions for lysine side-chain amino groups; and 2D refocused H(N)C and nonrefocused H(N.)C for arginine side-chain guanidino groups. These pulse sequences have been applied to perdeuterated 13C-/15N-labeled human carbonic anhydrase II (2H-HCA II). Because more than 95% of all side-chain 13C resonances in 2H-HCA II have already been assigned with the C(CC)(CO)NH experiment, the assignment of the side-chain 1HN/15N resonances has been straightforward using the pulse sequences mentioned above. The importance of assigning these side-chain HN protons has been demonstrated by recent studies in which the calculation of protein global folds was simulated using only 1HN-1HN NOE restraints. In these studies, the inclusion of NOE restraints to side-chain HN protons significantly improved the quality of the global fold that could be determined for a perdeuterated protein [R.A. Venters et al. (1995) J. Am. Chem. Soc., 117, 9592–9593].To whom correspondence should be addressed. 相似文献
12.
13.
Reactions of Cr(CO)3(η6-BT), in which the Cr is π-coordinated to the benzene ring of benzo[b]thiophene (BT), with Cp′(CO)2Re(THF), where Cp′ = η5-C5H5 or η5-C5Me5, give the products Cp′(CO)2Re(η2:η6-μ2-BT)Cr(CO)3 in which the Cr remains coordinated to the benzene ring and Re is bound to the C(2)=C(3) double bond. An X-ray diffraction study of Cp(CO)2Re(η2:η6-μ2-BT)Cr(CO)3 (3) provides details of the geometry. This structure contrasts with that of the Cp′(CO)2Re(BT) complexes that exist as mixtures of isomers in which the BT is coordinated to the Re through either the double bond (2,3-η2) or the sulfur (η1(S)). Thus, the electron-withdrawing Cr(CO)3 group in 3 stabilizes the 2,3-η2 mode of BT coordination to the Cp′(CO)2Re fragment. Implications of these results for catalytic hydrodesulfurization of BT are discussed. Crystal data for 3: triclinic, space group . 相似文献
14.
Body water content was determined on groups of Cyprinodon variegatus after sequential acclimation to ambient salinities in the range from fresh water to 100%. A multiple linear regression model applied to the data included body mass ( M ) of individuals and acclimation salinity ( S ) as independent variables, and body water content ( W ) as the dependent variable. The model that described the relationship was W (g) = 0·059 + 0·723 M (g)–0001 S (%). Converting values of body water content to percent of wet body mass produced a range of values from 74·5% in fresh water to 72·0% at 100%, only a 2·5% difference in body water content over this wide range of ambient salinities. 相似文献
15.
Tadashi II Masayuki Kubota Takashi Hirano Mamoru Ohashi Keiichi Yoshida Sakaru Suzuki 《Glycoconjugate journal》1995,12(3):282-289
The fast atom bombardment (FAB) collision induced dissociation (CID)-mass spectrometry/mass spectrometry (MS/MS) technique was successfully applied to characterize and identify the structures of the immunoreactive trisulfated and tetrasulfated tetrasaccharides that were obtained from the chondroitin sulfate in a shark fin using a treatment with chondroitinase ABC.Abbreviations FABMS
fast atom bombardment mass spectrometry
- CID
collision induced dissociation
- MS/MS
mass spectrometry/mass spectrometry
- UA2S-GalNAc6S
2-acetamido-2-deoxy-3-O-(2-O-sulfo--d-gluco-4-enepyranosyluronic acid)-6-O-sulfo-d-galactose
- UA-GalNAc4S
2-acetamido-2-deoxy-3-O-(-d-gluco-4-enepyranosyluronic acid)-4-O-sulfo-d-galactose
- UA-GalNAcDiS
2-acetamido-2-deoxy-3-O-(-d-gluco-4-enepyranosyluronic acid)-4,6-di-O-sulfo-d-galactose 相似文献
16.
Richard L. Rapport II George A. Ojemann Allen R. Wyler Arthur A. Ward Jr. 《The Western journal of medicine》1977,127(3):185-189
Therapy with anticonvulsant drugs reduces the frequency and severity of seizures in many but not all epileptic patients. Unfortunately, in a significant number control remains poor even when maximal doses of multiple anticonvulsant drugs are given. Some of these patients are candidates for surgical treatment of epilepsy. The operative management of convulsive disorders is a well-established technique and is available in some centers. In selected cases, such operations are both safe and effective, with good longterm improvement or complete control in 76 percent of patients. We have summarized the 24-year experience with surgical operation for epilepsy at the University of Washington Medical Center. 相似文献
17.
Smith P. R.; Ropelewski A. J.; Balog D.A.; Gottesman S.; Deerfield D. W. II 《Bioinformatics (Oxford, England)》1991,7(4):501-507
A method has been developed to link the display ability of ahigh-resolution graphics workstation with the computationalpower of a local mainframe or a remote supercomputer via anelectronic data network. The method allows this link to be establishedin a manner largely transparent to the user. The applicationof the method is illustrated by our successful distributionof the computationally intensive portions of an imaging program(MDPP) from a small VAX workstation to a VAX mainframe and CrayY-MP8/832 using a simple message-passing technique. This techniquecan be applied to almost any configuration of networked machines.
Received on February 5, 1991; accepted on April 23, 1991 相似文献
18.
Epstein-Barr Virus Polypeptides: Identification of Early Proteins and Their Synthesis and Glycosylation 总被引:6,自引:5,他引:1 下载免费PDF全文
We have identified at least six early polypeptides induced by Epstein-Barr virus in cells or under conditions which are nonpermissive for Epstein-Barr virus DNA replication ranging in molecular weight from 140,000 to 26,000. 相似文献
19.
WILLIAM F. KOCH II 《Lethaia: An International Journal of Palaeontology and Stratigraphy》1981,14(2):83-103
The Emsian? through early Eifelian Onondaga Limestone of the Appalachian Basin was deposited in a topographic basin and on the carbonate platform which surrounded the basin on the west, north, and northeast. Onondaga strata thin from the platform into the basin. Two sedimentary cycles are present in the sub-Tioga Onondaga of eastern North America. The Edgecliff-Amherstburg represents an interval of transgression, in which epeiric seas spread over much of eastern North America. During the Nedrow-Lucas regression, the interior of the carbonate platform became restricted, resulting in the deposition of evaporites. The Moorehouse-Anderdon transgression continued through the deposition of the Tioga Bentonite, followed by the pre-Speeds-Dundee regression from the craton. Early Eifelian Appalachian Basin Onondaga brachiopod communities, arranged from nearshore to offshore, include the Atrypid-Megakozlowskiella, Atrypid-Levenea, Chonetid, Atlanticocoelia, Ambocoeliid, and Truncalosia Communities. The Onondaga-age Eastern Americas Realm is divided into the Appohimchi Province in the Appalachian Basin and the Michigan Basin-Hudson Bay Lowland Province in the Midwest. The provincial assignment of the James Bay region of Ontario is uncertain; the Eastern Townships of Quebec are near the boundaries both of the two provinces of the Eastern Americas Realm, and of the Eastern Americas Realm and the Old World Realm, the latter realm being probably in the Canadian Maritime Provinces. 相似文献
20.
William H. Frey II Susan E. Senogles Leonard L. Heston Vicente B. Tuason Susan E. Nicol 《Journal of neurochemistry》1980,35(6):1418-1430
Abstract: Partial purification of soluble guanylate cyclase on DEAE-Sephacel yields two separate peaks of guanylate cyclase activity. After 10-fold purification of the soluble enzyme, guanylate cyclase is markedly inhibited by micromolar concentrations of dopamine (I50= 0.2 μm). Dopamine inhibition is observed whether the reaction is conducted with Mn21 or with Mg2+, under atmosphere or N2(g), and using enzyme from either peak from the DEAESephacel column. Other catecholamines also inhibit partially purified guanylate cyclase with an order of potency at 1 μm of: dopamine =l -DOPA > norepinephrine = isoproterenol = adrenochrome > epinephrine. The structural requirements for inhibition are two free hydroxyl groups on the phenyl ring and an ethylamine side chain. Dopamine also inhibits the Triton X-100-solubilized microsomal guanylate cyclase after partial purification on DEAESephacel. Neither chlorpromazine, propranolol, nor phentolamine at 20 μm effectively block the dopamine inhibition of partially purified soluble guanylate cyclase. Micromolar concentrations of the reducing agents dithiothreitol and glutathione also inhibit partially purified guanylate cyclase, but unlike these agents, catecholamines can inhibit whether added in the reduced or the oxidized forms. Inhibition of enzyme activity by micromolar concentrations of dopamine, adrenochrome, or dithiothreitol is rapidly reversed by dilution and the dopamine inhibition is competitive with MgGTP. Inhibition does not appear to involve covalent binding or to result from the ability of catecholamines to reduce the concentrations of oxygen or free radicals in solution. 相似文献