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301.
Esterase 1, a well-characterized mouse plasma protein of unknown function, has activity against a wide range of ester substrates including beta-alanine nitrophenyl esters and 17 beta-esters of estradiol. In this article, we report that esterase 1 is also responsible for a majority of the phorbol-12-ester hydrolase activity in mouse plasma. Incubation of homogeneous esterase 1 with 4 beta-phorbol 12 beta-myristate 13 alpha-acetate (PMA) at either 4 or 37 degrees C for up to 18 h yielded phorbol 13 alpha-acetate as the only hydrolysis product. Specific polyclonal antibodies to esterase 1 inhibited 95% of PMA hydrolysis by a purified esterase 1 preparation and 65% of PMA hydrolysis by mouse plasma. Perfused mouse liver homogenates contain two distinct phorbol diester hydrolases with apparent molecular masses of 65 kDa and 56 kDa, respectively. The 65-kDa protein appears to be immunologically identical to the plasma enzyme, while the 56-kDa protein, found in liver but not in plasma, is immunologically distinct. Phorbol 12-myristate, phorbol 12,13-dibutyrate, and PMA were found to be competitive inhibitors of the beta-alanine-nitrophenyl esterase activity of esterase 1 with Ki values of approximately 7 microM. Phorbol 13-acetate and phorbol itself were less effective with Ki values of 37 and 140 microM, respectively. Sodium salts of valeric and myristic acids did not inhibit at 10 microM. The above results indicate that efficient substrate binding requires a phorbol 12-ester. Similar results were obtained with estradiol 17 beta-valerate which is a better substrate for esterase 1 than is PMA. Our results strongly suggest that esterase 1 and a recently described phorbol ester hydrolase isolated from mouse serum (Saito, M., and Egawa, K. (1984) J. Biol. Chem. 259, 5821-5826) are the same and are immunologically and kinetically distinct from the 56-kDa phorbol 12-ester hydrolase in mouse liver. 相似文献
302.
G Fischer M Domingo D Lodder N Katz M Reinacher E Eigenbrodt 《Virchows Archiv. B, Cell pathology including molecular pathology》1987,53(6):359-364
Preneoplastic liver lesions were produced in female Wistar rats by application of 25 mg/kg N-nitrosomorpholine (NNM), 14 mg/kg diethylnitrosamine (DENA), 0.075 mg/kg aflatoxin B1 (AFB1) or 160 mg/kg safrole. These carcinogens were administered in two equal doses 12 and 24 h after partial hepatectomy. The animals then received sodium phenobarbital (0.1% in tap water) for up to 410 days. Numerous altered hepatic foci (AHF) and hyperplastic nodules (HN) were detected enzyme histochemically by their negative ATPase reaction after application of AFB1, DENA and NNM; some AHF and HN were also caused by the weak carcinogen safrole. Immunohistochemically these lesions were also L-pyruvate kinase (L-PK)-negative with a high coincidence with regard to their number and area. These results confirm the role of L-PK, an enzyme affecting the pentose phosphate pathway, as a negative marker of preneoplastic liver lesions. 相似文献
303.
304.
[1-14C] Xylitol was used as substrate for isolated liver parenchymal cells from fasted rats or hamsters, and the glucose formed was isolated and degraded. Over 90% of the total radioactivity was in carbons 1 and 3, and the ratio of specific activity of C-1 to that of C-3 was only slightly under two, the expected ratio for the classical mechanism of the non-oxidative pentose phosphate pathway. 相似文献
305.
306.
Sidney A. Katz 《Biological trace element research》1993,36(1):1-5
Various animal models have been employed for research on the significance of measuring trace element concentrations in the
human scalp hair. The objects of such research were the establishment of relationships between the concentrations of trace
elements in human scalp hair and (1) their concentrations in other compartments of the human body or (2) specific pathophysiological conditions. The guinea pig appears to be the animal of choice for such studies because the elemental
composition and growth pattern of its hair parallel those of the human scalp hair. 相似文献
307.
A histotopographic study of the nasal septum mucosa in rats was made using semi-serial sections stained with PAS-hematoxylin, reconstructed in form of maps representing the structure in a sagittal plane. The stratified squamous, respiratory and olfactory epithelia and Masera's organ cover 14.8, 43.6, 41.6 and 1.8%, respectively, of the septal surface (117.1 mm2). In the vestibular region, only ducts of PAS-negative glands of the respiratory region are found, and below the septum there is the infraseptal gland with PAS-negative acini. In the respiratory region, PAS-negative acinous glands form two groups: the superior and the inferior one occupying 10.5 and 1.5%, respectively, of the septal area. PAS-positive acinous glands are in the inferior half of the respiratory region and in a small anteroinferior portion of the olfactory region. Besides goblet cells broadly distributed, the respiratory epithelium presents scattered intraepithelial PAS-positive glands which are concentrated in the anterior portion and close to the nasopharyngeal duct. In the olfactory region prevail Bowman's PAS-positive glands which are also present in the mucosa of Masera's organ, but are not seen in the olfactory mucosa of Jacobson's organ. In the latter, PAS-positive glands are found in the respiratory mucosa. Globular leukocytes, cells of connective tissue origin, are constantly infiltrating the superior regions of the respiratory and olfactory epithelia, being more numerous in female rats. 相似文献
308.
309.
Homologous recombination and the repair of double-strand breaks during cotransformation of Dictyostelium discoideum. 总被引:2,自引:0,他引:2
We examined the ability of unlinked nonreplicating plasmid molecules to undergo homologous recombination during cotransformation of Dictyostelium amoebae. The transformation vector B10S confers resistance to the antibiotic G418 and was always presented to amoebae as a closed circle. Cotransforming DNA, containing a slime mold cDNA and sequences homologous to the primary vector, was presented either as a closed circle or as a linear molecule after digestion with restriction endonucleases which cut within one of three distinct regions of the plasmid. Remarkably, homologous recombination occurred in every clone examined. Moreover, the products of recombination were identical in all instances, irrespective of the presence or position of linearized ends. The ends of the linear templates were not recombinogenic. Repair of the introduced double-strand break occurred frequently during recombination. The repair could occur intermolecularly or, more likely, intramolecularly, i.e., by recircularization. Many of the recombination events were of a nonreciprocal nature. Despite the startlingly frequent level of homologous recombination, the use of cotransforming DNA which contains no homology to the selected vector established that such recombination was not required for cotransformation. 相似文献
310.
The effects of adenylyl methylene diphosphate (AMD), a non-hydrolyzable ATP analogue, were examined in sarcoplasmic reticulum vesicles isolated from rabbit skeletal muscle. The Ca2+-dependent APTase activity measured at 5 degrees C and pH 7.0 in 5.2 micrometer [gamma-32P]ATP and in the absence of added alkali metal salts was stimulated by added AMD. The steady state level of phosphoenzyme, however, was not decreased greatly by added AMP under these conditions. The hydrolysis of the phosphoenzyme formed at the steady state in the absence of added alkali metal salts was accelerated by added AMD to an extent that can account for the stimulation of the ATPase activity. At 5 degrees C and pH 7.0 the maximum stimulation of phosphoenzyme hydrolysis by AMD and the Km value for this ATP analogue were 4.3-fold and 40 micrometer, respectively. These results provide further support for our previous conclusion (Shigekawa, M., Dougherty, J.P. and Katz, A.M. (1978) J.Biol. Chem. 253, 1442--1450) that 2 classes of ATP site exist in the calcium pump ATPase in the absence of added alkali metal salts, one being the catalytic site and the other being the regulation site which activates the activity of the catalytic site. 相似文献