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11.
Specific chemical modifications of link protein and their effect on binding to hyaluronate and cartilage proteoglycan 总被引:6,自引:0,他引:6
M Lyon 《Biochimica et biophysica acta》1986,881(1):22-29
Specific chemical modifications of amino acid residues were performed on purified, native link protein from bovine articular cartilage. The effects of these on link protein's interactions with hyaluronate and bovine articular cartilage proteoglycan were assayed by gel chromatography. Interaction with hyaluronate was significantly perturbed by modification of lysine, arginine, tyrosine and aspartic/glutamic acid residues, but not histidine and tryptophan residues. No free, accessible sulphydryl group was found on native link protein. The requirement for unmodified lysine and arginine residues resembles that of the hyaluronate-binding site of pig laryngeal cartilage proteoglycan (Hardingham, T.E., Ewins, R.J.F. and Muir, H. (1976) Biochem. J. 157, 127-143). In contrast, proteoglycan binding was only significantly perturbed by the loss of arginine residues. This resistance may reflect hydrophobicity of the binding site or masking of the site from chemical modification by link protein self-association. Amidation of carboxyl groups, which destroyed hyaluronate binding but left proteoglycan binding intact, provides a means of generating a monofunctional link protein molecule of potential use in proteoglycan aggregation studies. 相似文献
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The soil isolate Pseudomonas putida MW1000 can grow on toluene and other hydrocarbons; in this respect it is similar to strains of Pseudomonas which carry the TOL plasmid. By conjugation experiments, the genes conferring these growth abilities have been shown to be located on the bacterial chromosome, linked to vil and catB. A 56-kilobase segment of the bacterial chromosome of MW strains carrying the TOL genes can transpose to the IncP-1 plasmid R18-18. Physical analysis of these TOL R18-18 hybrids has shown that the TOL segment is almost identical to the same region found in the TOL plasmid pWW0. 相似文献
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In order to evaluate whether irradiation of the postimplantation maternal environment contributed to the induction of postimplantation mortality or congenital anomalies, mouse ovaries were surgically exteriorised and selectively irradiated or shielded in a specially constructed apparatus. The results show that exposure of the mouse abdomen and uterus to 3.70 Gy X-rays, 15-21 days prior to conception, has no significant effect on the incidence of either postimplantation mortality or congenital anomalies. Exposure of the ovaries to 3.27 Gy X-rays during the same period, however, increased the frequency of both postimplantation mortality and congenital anomalies. 相似文献
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The induction of congenital malformations among the offspring of male mice treated with X-rays at pre-meiotic and post-meiotic stages has been studied in two experiments. Firstly, animals were exposed to varying doses (108–504 cGy) of X-rays and mated at various time intervals (1–7, 8–14, 15–21 and 64–80 days post-irradiation), so as to sample spermatozoa, spermatids and spermatogonial stem cells. In the second experiment, only treated spermatogonial stem cells were sampled. One group of males was given a single 500-cGy dose, a second group a fractionated dose (500 + 500 cGy, 24 h apart) and a third group was left unexposed.In the first experiment, induced post-implantation dominant lethality increased with dose, and was highest in week 3, in line with the known greater radiosensitivity of the early spermatid stage. Preimplantation loss also increased with dose and was highest in week 3. There was no clear induction of either pre-implantation or post-implantation loss at spermatogonial stem cell stages.There was a clear induction of congenital malformations at post-meiotic stages, the overall incidence being 2.0 ± 0.32% in the irradiated series and 0.24 ± 0.17% among the controls. The induction was statistically significant at each dose. At the two highest doses the early spermatids (15–21 days) appeared more sensitive than spermatozoa, and at this stage the incidence of malformations increased with dose. The data from Expt. 1 on the induction of malformations by irradiation of spermatogonial stages were equivocal. In contrast, Expt. 2 showed a statistically significant induction of malformations at both dose levels (2.2 ± 0.46% after 500 cGy and 3.1 ± 0.57% after 500 + 500 cGy). The relative sensitivities of male stem cells, post-neiotic stages and mature oocytes to the induction of congenital malformations were reasonably similar to their sensitivities for specific-locus mutations, except that the expected enhancing effect of the fractionation regime used was not seen.Dwarfism and exencephaly were the two most commonly observed malformations in all series. 相似文献
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The relationship between cartilage thickness and proteoglycan extractability was examined. Bovine nasal cartilage slices (20, 100, and 500 micron thicknesses) were extracted with low-ionic-strength buffer and 4 M guanidine hydrochloride. The extractability of proteoglycans with both solutions depended on slice thickness. Thinner slices yielded greater amounts of proteoglycans. Sixty-three percent of the total cartilage uronic acid was extracted from 20-micron cartilage slices with low-ionic-strength buffer while only 7% was extracted for 500-micron slices. Each fivefold increase in cartilage surface area led to a threefold increase in uronic acid extraction with low-ionic-strength buffer. Extraction of proteoglycan aggregates was directly proportional to the cartilage surface area whereas extraction of non-aggregated proteoglycans, per surface area, increased with increasing cartilage thickness. These data are consistent with the hypothesis that proteoglycan aggregates are extracted mainly from the cartilage surface while non-aggregated proteoglycans diffuse from deep within the cartilage. Extraction with low-ionic-strength buffer occurred in two phases. There was an initial rapid loss of proteoglycans in which 1/3 to 1/2 of all proteoglycans eluting over 6 days were extracted during the first 30 min. Subsequent extraction was much slower with decreasing amounts extracted on each consecutive day. The initial rapid loss of proteoglycans was probably due to the steep osmotic-pressure gradient existing when the cartilage was placed in the low-ionic-strength buffer. 相似文献
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Mary F. Lyon 《Genetics》1974,78(1):305-309