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41.
Kleijer W. J. Huijmans J. G. M. Blom W. Gorska D. Kubalska J. Walasek M. Zaremba J. 《Human genetics》1984,66(4):287-288
Summary The prenatal diagnosis of a fetus affected with Sanfilippo disease type B is described. The deficiency of -N-acetylglucosaminidase in the cultured amniotic fluid cells was shown by a microassay enabling early prenatal diagnosis. In addition an increased level of heparan sulphate was demonstrated in the amniotic fluid by two-dimensional electrophoresis of glycosaminoglycans. The latter result confirmed the value of this test as an adjunctive method in the prenatal diagnosis. The pregnancy was terminated and the prenatal diagnosis was confirmed by enzyme analysis of cultured fetal fibroblasts and fetal liver. 相似文献
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Mindaugas Zaremba Amelia Owsicka Gintautas Tamulaitis Giedrius Sasnauskas Luda S. Shlyakhtenko Alexander Y. Lushnikov Yuri L. Lyubchenko Niels Laurens Bram van den Broek Gijs J. L. Wuite Virginijus Siksnys 《Nucleic acids research》2010,38(20):7142-7154
To cut DNA at their target sites, restriction enzymes assemble into different oligomeric structures. The Ecl18kI endonuclease in the crystal is arranged as a tetramer made of two dimers each bound to a DNA copy. However, free in solution Ecl18kI is a dimer. To find out whether the Ecl18kI dimer or tetramer represents the functionally important assembly, we generated mutants aimed at disrupting the putative dimer–dimer interface and analysed the functional properties of Ecl18kI and mutant variants. We show by atomic force microscopy that on two-site DNA, Ecl18kI loops out an intervening DNA fragment and forms a tetramer. Using the tethered particle motion technique, we demonstrate that in solution DNA looping is highly dynamic and involves a transient interaction between the two DNA-bound dimers. Furthermore, we show that Ecl18kI cleaves DNA in the synaptic complex much faster than when acting on a single recognition site. Contrary to Ecl18kI, the tetramerization interface mutant R174A binds DNA as a dimer, shows no DNA looping and is virtually inactive. We conclude that Ecl18kI follows the association model for the synaptic complex assembly in which it binds to the target site as a dimer and then associates into a transient tetrameric form to accomplish the cleavage reaction. 相似文献
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The BfiI endonuclease cleaves DNA at fixed positions downstream of an asymmetric sequence. Unlike other restriction enzymes, it functions without metal ions. The N-terminal half of BfiI is similar to Nuc, an EDTA-resistant nuclease from Salmonella typhimurium that belongs to the phosphoplipase D superfamily. Nuc is a dimer with one active site at its subunit interface, as is BfiI, but it cuts DNA non-specifically. BfiI was cleaved by thermolysin into an N-terminal domain, which forms a dimer with non-specific nuclease activity, and a C-terminal domain, which lacks catalytic activity but binds specifically to the recognition sequence as a monomer. On denaturation with guanidinium, BfiI underwent two unfolding transitions: one at a relatively low concentration of guanidinium, to a dimeric non-specific nuclease; a second at a higher concentration, to an inactive monomer. The isolated C-terminal domain unfolded at the first (relatively low) concentration, the isolated N-terminal at the second. Hence, BfiI consists of two physically separate domains, with catalytic and dimerisation functions in the N terminus and DNA recognition functions in the C terminus. It is the first example of a restriction enzyme generated by the evolutionary fusion of a DNA recognition domain to a phosphodiesterase from the phospholipase D superfamily. BfiI may consist of three structural units: a stable central core with the active site, made from two copies of the N-terminal domain, flanked by relatively unstable C-terminal domains, that each bind a copy of the recognition sequence. 相似文献
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W S Simonides R Zaremba C van Hardeveld W J van der Laarse 《Analytical biochemistry》1988,169(2):268-273
A method for the determination of lactate is described in which the esterification of lactate with the uv-absorbing compound alpha-p-dibromoacetophenone is followed by separation and quantitation of the ester by reversed-phase HPLC with detection at 254 nm. The reproducibility, detection limit, and precision of the method are comparable to those of conventional methods which use enzymatic cycling for enhanced performance. The applicability of this rapid and simple method is illustrated with the determination of picomole amounts of lactate in resting and stimulated single muscle fibers. 相似文献
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Danguole Satkunskiene Tiago M. da Silva Sigitas Kamandulis Nuno M.C. Leite Aurelijus Domeika Mantas Mickevicius Audrius Snieckus 《Journal of musculoskeletal & neuronal interactions》2020,20(4):488
Objective:the purpose of this study was to identify differences in hamstring passive stiffness between the pre-season and in-season periods.Methods:Hamstring strength and passive stiffness were measured in professional male soccer players before and after the pre-season (4 weeks), and after the in-season (6 weeks) periods using an isokinetic dynamometer. Muscle passive stiffness was determined from the slope of the passive torque–angle relationship. External loads (acceleration and jumps) were monitored by GPS and internal loads by questionnaire.Results:Hamstring passive stiffness increased after 10 weeks of training and matches, without changes in passive peak torque and range of motion. The hamstring passive stiffness modifications were associated with the volume and intensity of accelerations and jumps. The individual data analysis also provided some support for the suppression of the biomechanical adaptation in the subjects with relatively large external load.Conclusions:Regular training and match workouts increase hamstring passive stiffness in professional soccer players but the adaptation of muscle-tendon unit passive elements might not occur if players experience excessive mechanical stress. 相似文献