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41.
Maurizio Pellecchia Gerhard Wider Hideo Iwai Kurt Wüthrich 《Journal of biomolecular NMR》1997,10(2):193-197
A novel 2D NMR experiment, 2D HE(NE)HGHH, is presented for the assignment ofarginine side chain 1H and 15N resonances inuniformly 15N-labeled proteins. Correlations between1H, 1Hand 1H are established on the basis of3J(15N,1H) heteronuclear scalarcoupling constants, and sequence-specific assignments are obtained by overlapof these fragments with 1H chemical shiftsobtained by assignment procedures starting from the polypeptide backbone.Since guanidino protons exchange quite rapidly with the bulk water, the 2DHE(NE)HGHH pulse scheme has been optimized to avoid saturation and dephasingof the water magnetization during the course of the experiment. As anillustration, arginine side chain assignments are presented for two uniformly15N-labeled proteins of 7 and 23 kDa molecular weight. 相似文献
42.
S. Aime Mauro Botta S. Geninatti Crich Giovanni B. Giovenzana Roberto Pagliarin Maurizio Piccinini Massimo Sisti Enzo Terreno 《Journal of biological inorganic chemistry》1997,2(4):470-479
A novel heptacoordinating ligand consisting of a thirteen-membered tetraazamacrocycle containing the pyridine ring and bearing
three methylenephosphonate groups (PCTP-[13]) has been synthesized. Its Gd(III) complex displays a remarkably high longitudinal
water proton relaxivity (7.7 mM–1 s–1 at 25 °C, 20 MHz and pH 7.5) which has been accounted for in terms of contributions arising from (1) one water molecule
bound to the metal ion, (2) hydrogen-bonded water molecules in the second coordination sphere, or (3) water molecules diffusing
near the paramagnetic chelate. Variable-temperature 17O-NMR transverse relaxation data indicate that the residence lifetime of the metal-bound water molecule is very short (8.0 ns
at 25 °C) with respect to the Gd(III) complexes currently considered as contrast agents for magnetic resonance imaging. Furthermore,
GdPCTP-[13] interacts with human serum albumin (HSA), likely through electrostatic forces. By comparing water proton relaxivity
data for the GdPCTP-[13]-HSA adduct, measured as a function of temperature and magnetic field strength, with those for the
analogous adduct with GdDOTP (a twelve-membered tetraaza macrocyclic tetramethylenephosphonate complex lacking a metal-bound
water molecule), it has been possible to propose a general picture accounting for the main determinants of the relaxation
enhancement observed when a paramagnetic Gd(III) complex is bound to HSA. Basically, the relaxation enhancement in these systems
arises from (1) water molecules in the hydration shell of the macromolecule and protein exchangeable protons which lie close
to the interaction site of the paramagnetic complex and (2) the metal bound water molecule(s). As far as the latter contribution
is concerned, the interaction with the protein causes an elongation of the residence lifetime of the metal-bound water molecule,
which limits, to some extent, the potential relaxivity enhancement expected upon the binding of the paramagnetic complex to
HSA.
Received: 27 January 1997 / Accepted: 12 May 1997 相似文献
43.
CD9, a tetraspan transmembrane protein, renders cells susceptible to canine distemper virus. 总被引:13,自引:3,他引:10 下载免费PDF全文
S Lffler F Lottspeich F Lanza D O Azorsa V ter Meulen J Schneider-Schaulies 《Journal of virology》1997,71(1):42-49
Canine distemper virus (CDV), a lymphotropic and neurotropic negative-stranded RNA virus of the Morbillivirus genus, causes a life-threatening disease in several carnivores, including domestic dogs. To identify the cellular receptor(s) involved in the uptake of CDV by susceptible cells, we isolated a monoclonal antibody (MAb K41) which binds to the cell surface and inhibits the CDV infection of several cell lines from various species. Pretreatment of cells with MAb K41 reduces the number of infectious centers and the size of the syncytia. Using affinity chromatography with MAb K41, we purified from HeLa and Vero cell extracts a 26-kDa protein which contained the amino acid sequence TKDEPQRETLK of human CD9, a member of the tetraspan transmembrane or transmembrane 4 superfamily of cell surface proteins. Transfection of NIH 3T3 or MDBK cells with a CD9 expression plasmid rendered these cells permissive for viral infection and raised virus production by a factor of 10 to 100. The mechanism involved is still unclear, since we were unable to detect direct binding of CDV to CD9 by using immunoprecipitation and a virus overlay protein binding assay. These findings indicate that human CD9 and its homologs in other species are necessary factors for the uptake of CDV by target cells, the formation of syncytia, and the production of progeny virus. 相似文献
44.
Maurizio Meta Marco Ponte Marina Guastella Claudia Semino Gabriella Pietra Giovanni Battista Ratto Giovanni Melioli 《Cancer immunology, immunotherapy : CII》1995,40(4):235-240
Despite the combined use of surgery and chemoradiotherapy, the poor prognosis of advanced non-smallcell lung cancer (NSCLC) requires the definition of new therapeutic approaches. The presence of T lymphocytes, with peculiar phenotypic, functional and molecular characteristics within the tumour, suggested the possible use of these cells, expanded in vitro, in protocols of adoptive immunotherapy. We have described how a population of oligoclonal T lymphocytes, derived from advanced NSCLC, can be expanded in vitro and has the capability of lysing autologous cancer cells. What is more important, we observed that patients with advanced NSCLC, treated with TIL expanded in vitro and recombinant interleukin-2, seemed to have a disease-free period longer than that of patients treated with conventional chemoradiotherapy. in an attempt to find new sources of specific lymphocytes for immunotherapy, we describe the analysis of the phenotypic, functional and molecular characteristics of T lymphocytes, derived from lymph nodes draining advanced NSCLC. In this paper we show that these cells, have restriction patterns of T cell receptor chain similar to those detectable in the population of infiltrating T lymphocytes. This finding suggests that T cells derived from draining lymph nodes of advanced NSCLC have peculiar characteristics and can be a suitable source of effector cells for protocols of adoptive immunotherapy in lung cancer treatment. 相似文献
45.
Maurizio Zandomeneghi 《Chirality》1995,7(6):446-451
Complexation of 2-(3′-benzoylphenyl)propionic acid (ketoprofen), 1 , to bovine serum albumin (BSA) results in an intense negative circular dichroism in the ketonic n → π* band of the benzoylphenyl moiety. This high CD contrasts with the weak CD of 1 -enantiomers dissolved in common solvents. Furthermore, a number of chiral and achiral molecules containing the benzophenone moiety are easily complexed to BSA: all these complexes show an intense CD at the same transition. To account for the observed CD intensities of the above molecules, it appears that BSA complexation markedly shifts the equilibrium between strongly asymmetric, antipodic conformers. Dissymmetry of these conformers is connected to the instability of a structure with phenyl rings coplanar to the carbonyl chromophore, as also indicated by molecular mechanics calculations. The magnification of the Cotton effects of the 1 -antipodes, due to the protein, can be used to measure the optical purity of 1 -samples with excellent precision. In contrast with BSA, human SA is unable to recognize the chirality of 1 -antipodes; oleic acid cocomplexation modifies this fact as well as other features of the binding. © 1995 Wiley-Liss, Inc. 相似文献
46.
Maurizio Pacifici 《Matrix biology》1995,14(9)
In comparison to the vast literature on articular cartilage structure and function, relatively little is known about how articular cartilage forms during embryo-genesis and is endowed with unique phenotypic properties, most notably the ability to persist and function throughout postnatal life. In this minireview, we summarize recent studies from our laboratory suggesting that the extracellular matrix protein tenascin-C is involved in the genesis and function of articular chondrocytes. These and other data have led us to propose that tenascin-C may be part of in vivo mechanisms whereby articular chondrocytes develop at the epiphysis of long bone models, remain functional throughout postnatal life, and avoid the endochondral ossification process undertaken by the bulk of chondrocytes located in the metaphysis and diaphysis of skeletal models. 相似文献
47.
Asellus aquaticus, a sexually dimorphic isopod crustacean, has no morphologically distinguishable sex chromosome pair. In 25% of the males from a natural population the fluorochrome chromomycin A3 revealed a pair of metacentric chromosomes that was heteromorphic for two heterochromatic areas. One chromosome had a fluorescent band in each of the two arms, whereas the other chromosome lacked both bands. Chromosomal analysis was performed on 50 sibships produced by known parents. The heterochromosome was inherited only by the male offspring of males bearing it. Its distribution led us to postulate the existence of an incipient differentiating sex chromosome pair in the A. aquaticus population.This paper is dedicated to Prof. Guiseppe Montalenti on the occasion of his 80th birthday. 相似文献
48.
Leilo Zolla Harry A. Kuiper Alessandro Finazzi Agrò Maurizio Brunori 《Journal of inorganic biochemistry》1984,22(3):143-153
Addition of KCN to Helix pomatia β-hemocyanin fully saturated with either O2 or CO results in a decrease of the spectroscopic properties of the protein (absorbance at 340 nm and luminescence at 550 nm) due to the displacement of the gaseous ligands (O2 or CO) from the active site. The anionic form of cyanide (CN?) is supposed to bind to the active site; its intrinsic affinity for the protein, as calculated from independent O2 and CO displacement experiments, is between 2 and 6 × 106M?1. The replacement of O2 or CO shows some differences which may be correlated with the different modes of binding at the active site. Thus, while displacement of oxygen by cyanide is hyperbolic, addition of cyanide to carbonylated hemocyanin shows a lag phase. This finding suggests the formation of a mixed liganded complex at the active site. The simultaneous presence of CO and CN? at the active site of hemocyanin is also supported by the experiment in which addition of small amounts of KCN to hemocyanin partially saturated with O2 and CO gives rise to an increase of emission intensity and a concomitant decrease of the O2 absorption band. The mixed-liganded species displays luminescence properties similar to those of CO-saturated hemocyanin, and the formation of the complex is reversible on dialysis or oxygenation. 相似文献
49.
50.
During germination, the transmembrane electric potential (PD) of cortical cells of the embryonal axis of radish seeds (Raphanus sativus L.) rises from-120 mV initially to a maximum of-150 mV after 5 h incubation, then falls again to stable values of around-120 mV. Treatments inhibiting germination block the transitory PD increase. Administration of uncoupling agents or low temperatures, during the process of germination, produces a marked fall of the PD transitory increase. Abscisic Acid has a parallel inhibitory effect on PD and germination, while fusicoccin produces a rise in both; administration of abscisic acid with fusicoccin inhibits germination, while the PD remains at the high levels given by fusicoccin. These results are discussed in relation to ion exchange at membrane level.Abbreviations ABA
abscisic acid
- FC
fusicoccin
- GA3
gibberellic acid
- PD
electric potential difference (between the vacuole and the external medium)
- CH
cycloheximide
- DNP
dinitrophenol
- FCCP
(p-trifluormethoxy)-carbonylcyanide-phenylhydrazone
- DCCD
N,N-dicyclohexylcarbodiimide 相似文献