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111.
In a previous report, the cDNA for human proteinase inhibitor 8 (PI8) was first identified, isolated, and subcloned into a mammalian expression vector and expressed in baby hamster kidney cells. Initial studies indicated that PI8 was able to inhibit the amidolytic activity of trypsin and form an SDS-stable approximately 67-kDa complex with human thrombin [Sprecher, C. A., et al. (1995) J. Biol Chem. 270, 29854-29861]. In the present study, we have expressed recombinant PI8 in the methylotropic yeast Pichia pastoris, purified the inhibitor to homogeneity, and investigated its ability to inhibit a variety of proteinases. PI8 inhibited the amidolytic activities of porcine trypsin, human thrombin, human coagulation factor Xa, and the Bacillus subtilis dibasic endoproteinase subtilisin A through different mechanisms but failed to inhibit the Staphylococcus aureus endoproteinase Glu-C. PI8 inhibited trypsin in a purely competitive manner, with an equilibrium inhibition constant (Ki) of less than 3.8 nM. The interaction between PI8 and thrombin occurred with a second-order association rate constant (kassoc) of 1.0 x 10(5) M-1 s-1 and a Ki of 350 pM. A slow-binding kinetics approach was used to determine the kinetic constants for the interactions of PI8 with factor Xa and subtilisin A. PI8 inhibited factor Xa via a two-step mechanism with a kassoc of 7.5 x 10(4) M-1 s-1 and an overall Ki of 272 pM. PI8 was a potent inhibitor of subtilisin A via a single-step mechanism with a kassoc of 1.16 x 10(6) M-1 s-1 and an overall Ki of 8.4 pM. The interaction between PI8 and subtilisin A may be of physiological significance, since subtilisin A is an evolutionary precursor to the intracellular mammalian dibasic processing endoproteinases. 相似文献
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1. We examined the role of flooding on the leaf nutrient content of riparian trees by comparing the carbon : nitrogen : phosphorus (C : N : P) ratio of leaves and litter of Rio Grande cottonwood (Populus deltoides ssp. wislizenii) in flood and non‐flood sites along the Middle Rio Grande, NM, U.S.A. The leaf C : N : P ratio was also examined for two non‐native trees, saltcedar (Tamarix chinensis) and Russian olive (Elaeagnus angustifolia), and six species of dominant riparian arthropods. 2. Living leaves and leaf litter of cottonwoods at flood sites had a significantly lower leaf N : P ratio and higher %P compared with leaves and litter at non‐flood sites. A non‐flood site downstream from wastewater effluent had a significantly lower litter C : N ratio than all other sites, suggesting N fertilisation through ground water. The non‐native trees, saltcedar and Russian olive, had higher mean leaf N content, N : P ratio, and lower C : N ratio compared with cottonwoods across study sites. 3. Riparian arthropods ranged from 5.2 to 7.1 for C : N ratio, 56–216 for C : P ratio, and 8.9–34 for N : P ratio. C content ranged from 25 to 52% of dry mass, N content from 4.7 to 10.8%, and P content from 0.59 to 1.2%. Differences in stoichiometry between high C : nutrient leaf litter and low C : nutrient invertebrates suggests possible food‐quality constraints for detritivores. 4. These results suggest that spatial and temporal variation in the C : N : P ratio of cottonwood leaves and leaf litter is influenced by surface and subsurface hydrologic connection within the floodplain. Reach‐scale variation in the elemental composition of riparian organic matter inputs may have important implications for decomposition, nutrient cycling, and food webs in river floodplain systems. 相似文献
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SYNOPSIS. Supraspecific taxa of the ciliate Order Tintinnida Corliss, 1955 are indexed by 304 entries; genetic type species are cited with reference to the original citation. The accompanying bibliography of 133 entries includes those publications cited in the index. Authors for several type citations are corrected, as is authorship of some genera, and a new type designation is made for Parundella to replace the previous erroneous designation. Genera present in the fossil record are so indicated. 相似文献
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Daily studies were made of the osmoregulatory abilities of Cardisomaguanhumi from hatching to the end of larval development. Theseresults were compared to those of similar research on osmoregulationby larvae and megalopa of four species of estuarine, littoral,and sub-littoral crabs. The comparison shows that larvae ofC. guanhumi possess the same kinds of adaptations for waterintake at the time of molt that were found in Rhithropanopeusharrisii. Land-crab larvae hyperregulated in 10 p.p.t. sea waterand hyporegulated in water of 40 p.p.t. for experimental periodsfor 2 hr during the first one-third of their development. Duringthe remainder of larval life, they hyporegulated against 15p.p.t. in intermolt periods and became isosmotic with, or hyporegulatedagainst, 10 p.p.t. at the time of molting. From the time ofhatching, the osmoregulatory pattern of developing C. guanhumifits them for deep penetration of estuaries and for crossingsteep saline gradients. This pattern is evidence for a strongerand more enduring control of water balance, especially at thetime of molting, than we have found in non-terrestrial species. 相似文献
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Larvae of the land-crab, Cardisoma guanhumi, Latreille. weremaintained in 24 different combinations of salinity and temperaturefrom the time of hatching. Survival to the first crab occurredin salinities of 1545 p.p.t., 25° and 30°C. Durationof the five zoeal and one megalops stages was similar in salinitiesof 2040 p.p.t., but at 15 and 45 p.p.t. a greater periodof time was required for total development. Mortality of allthe larvae at 20°C suggests that temperature plays a moreimportant role in survival and distribution of the larvae ofC. guanhumi than salinity. Increments of size in crabs during the first seven post-larvalmolts were similar in salinities of 535 p.p.t., 25°C,but in fresh water increase in size at the time of molting wasreduced. Although there was no apparent relationship betweenfrequency of molting and salinities of 535 p.p.t., theduration of intermolt was reduced in crabs maintained in freshwater, and survival was also lower. From the present study there is no indication that the morphologicaland physiological processes that are associated with adaptationof the adult crab to the terrestrial environment are initiatedduring larval development. Although the adult crabs have successfullypenetrated the terrestrial environment, the pelagic larvae arestill subject to the numerous ecological variables of the estuarineand marine environments. 相似文献
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