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51.
Sharon X. Chen Charles C. Hardin Harold E. Swaisgood 《Journal of Protein Chemistry》1993,12(5):613-625
Incubation of -lactoglobulin with immobilized trypsin at 5–10°C results in a time-dependent release of several fragments of the core domain in yields approaching 15%. Digests were fractionated by ion-exchange chromatography with a Mono Q HR5/5 column and analyzed after disulfide reduction by polyacrylamide gel electrophoresis in sodium dodecylsulfate. Three fragments with approximate molecular weights of 13.8, 9.6, and 6.7 kD were identified. The fraction from ion-exchange chromatography yielding the 6.7 kD fraction after disulfide reduction was further characterized because it was most homogeneous and gave the highest yield. The C-terminal cleavage site of the 6.7 kD core fragment appeared to be Lys100 or Lys101 as determined by C-terminal amino acid analysis. The exact masses, after reduction with dithiothreitol, are 6195 and 6926 as determined by laser desorption mass spectrometry, corresponding to residues 48–101 and 41–100. Prior to reduction, -lactoglobulin C-terminal residues 149–162 are connected to these core domain fragments as shown by C-terminal analysis and mass spectrometry. Structural studies indicate that these 7.9 and 8.6 kD core domain fragments released by immobilized trypsin retain much of their native structure. CD spectra indicate the presence of antiparallel -sheet structure similar to the native protein but the -helix is lost. Spectra in the aromatic region indicate the existence of tertiary structure. Moreover, structural transitions in urea are completely reversible as measured by CD spectra, although the extrapolated G
D
H20
and the urea concentration at the transition midpoint are lower than for the native protein. The core domain fragments also display apH-dependent binding to immobilizedtrans-retinal as does intact protein. A single endotherm is obtained for both core domain fragments and native protein upon differential scanning calorimetry, but again, the domain is less stable as indicated by a transition peak maxima of 56.9°C as compared with 81.1°C for native protein.Abbreviations used: CD, circular dichroism; CPG, controlled pore glass; DSC, differential scanning calorimetry; DTT, dithiothreitol; FPLC, fast flow liquid chromatography; HPLC, high-performance liquid chromatography; PITC, phenylisothiocyanate; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; TEA, triethylamine; UV, ultraviolet. 相似文献
52.
Sharon A. Robinson Catherine E. Lovelock C. B. Osmond 《Plant biology (Stuttgart, Germany)》1993,106(4):307-312
The leaves of the Crassulacean acid metabolism plant Cotyledon orbiculata have a waxy coating which is highly reflective but can be easily removed by brushing. This provided an ideal system in which to investigate the role of epidermal wax as a possible photoprotectant. Removal of the wax, prior to exposure to natural sunlight, resulted in substantial decreases in Fv/Fm and in severe cases evidence of photoinhibitory damage, as indicated by a rise in Fo. Leaves from which wax had been removed also showed higher conversion of violaxanthin to zeaxanthin than waxed leaves. Recovery of brushed leaves over a 12 day period was correlated with an increase in the total pool of xanthophyll cycle components. This study suggests that the presence of highly reflective wax on the epidermis may confer significant photoprotection to plants exposed to high solar radiation environments. 相似文献
53.
Mutants in dolichol synthesis: conversion of polyprenol to dolichol appears to be a rate-limiting step in dolichol synthesis 总被引:3,自引:1,他引:2
Rosenwald Anne G.; Stanley Pamela; McLachlan Karen R.; Krag Sharon S. 《Glycobiology》1993,3(5):481-488
Chinese hamster ovary (CHO) cells of the Lec9 recessive complementationgroup display a distinctive profile of resistance to a varietyof toxic lectins. In addition, they accumulate cis--unsaturatedpolyprenol and use mainly polyprenol rather than dolichol tosynthesize the glycosylated lipids used in asparagine-linkedglycosylation of proteins. The primary defect in these cellsis thought to result from a deficiency in polyprenol reductaseactivity. Three new mutants were isolated and determined tohave qualitatively, although not quantitatively, similar lectinresistance profiles to Lec9 cells. Two of these mutants (AbrRand RicR) also contained polyprenol rather than dolichol. Thelectin resistance profile of an independent mutant which accumulatespolyprenol, F2A8, was also found to be qualitatively similarto the Lec9 pattern. The relationship among these mutants wasanalysed in more detail by construction of cellcell hybrids.Lectin resistance profiles of the hybrids demonstrated thatAbrR, RicR and F2A8 fell into the Lec9 complementation group.Analysis of prenols in the hybrids also showed that F2A8 wasa member of the Lec9 group. Surprisingly, a significant fractionof the prenols found in Lec9 Parent hybrids was polyprenol(up to 30% of the neutral fraction), whereas the prenols foundin Parent Parent hybrids were nearly exclusively dolichol(97% of the neutral lipid fraction). Therefore, reduction ofpolyprenol to dolichol appears to be a rate-limiting step inthe synthesis of dolichol since hybrids with differing numbersof wild-type alleles can be biochemically distinguished. CHO cells dolichol lectins mutants polyprenol reductase 相似文献
54.
55.
Barindra Sana Ke Ding Jia Wei Siau Rupali Reddy Pasula Sharon Chee Sharad Kharel Jean-Baptise Henri Lena Eunice Goh Lakshminarayanan Rajamani Yeng Ming Lam Sierin Lim John F. Ghadessy 《Biotechnology and bioengineering》2023,120(11):3200-3209
Polyethylene terephthalate (PET) hydrolase enzymes show promise for enzymatic PET degradation and green recycling of single-use PET vessels representing a major source of global pollution. Their full potential can be unlocked with enzyme engineering to render activities on recalcitrant PET substrates commensurate with cost-effective recycling at scale. Thermostability is a highly desirable property in industrial enzymes, often imparting increased robustness and significantly reducing quantities required. To date, most engineered PET hydrolases show improved thermostability over their parental enzymes. Here, we report engineered thermostable variants of Ideonella sakaiensis PET hydrolase enzyme (IsPETase) developed using two scaffolding strategies. The first employed SpyCatcher-SpyTag technology to covalently cyclize IsPETase, resulting in increased thermostability that was concomitant with reduced turnover of PET substrates compared to native IsPETase. The second approach using a GFP-nanobody fusion protein (vGFP) as a scaffold yielded a construct with a melting temperature of 80°C. This was further increased to 85°C when a thermostable PETase variant (FAST PETase) was scaffolded into vGFP, the highest reported so far for an engineered PET hydrolase derived from IsPETase. Thermostability enhancement using the vGFP scaffold did not compromise activity on PET compared to IsPETase. These contrasting results highlight potential topological and dynamic constraints imposed by scaffold choice as determinants of enzyme activity. 相似文献
56.
Kinley Ragan Jan Schipper Heather L. Bateman Sharon J. Hall 《The Journal of wildlife management》2023,87(1):e22322
Wildlife populations in semi-arid regions are increasingly challenged by human activities and dependent on the connectivity of riparian corridors for access to surface water. The Madrean Archipelago is a biodiversity hotspot along the arid United States–Mexico borderlands that support both Neotropical and Nearctic wildlife. Infrastructure development (e.g., the border wall and the expansion of Mexican Federal Highway 2) in this region inhibits wildlife movement along the transnational mountain archipelago by disconnecting habitat. To explore the relationship between habitat variables and mammal use of riparian corridors in northern Sonora, Mexico, we collected data from 19 motion-sensitive cameras between October 2018 and April 2019 and used single-season occupancy models and Royle-Nichols abundance estimation models to analyze our data. We recorded 21 species of mammals, including the first sighting of jaguar (Panthera onca) in this region in 25 years. River characteristics (distance from river, riparian corridor width, water availability), remoteness (distance from highway, productivity, elevation), and topographic variety (vertical elevation difference) influenced patterns of occupancy probability and estimated abundance of mammals >1 kg, but the strength and direction of these relationships varied by species. Additionally, intermittently wet desert washes were comparable in species richness to the perennial system. These results highlight the importance of examining physical and biological aspects of habitat. This is especially true when identifying corridors where mitigation structures should be placed to improve wildlife connectivity in biodiversity hotspots like the Madrean Archipelago and semi-arid ecosystems worldwide. 相似文献
57.
The aims of this study were to investigate the diet and relative abundance of fruit bats in a lowland Malaysian rain forest and to test the hypothesis that the local assemblage structure of fruit bats varies significantly over time in relation to the availability of food. In total, 352 fruit bats of eight species were captured during 72,306 m2 mist‐net hours of sampling between February 1996 and September 1999. Three species of fruit bats (Balionycteris maculita, Chironax melanocephalus, and Cynopterus brachyotis) that fed on a wide range of “steady state” and “big bang” food resources were captured continuously throughout the study period, with no significant variation in capture rates over time. In contrast, five species that fed exclusively or almost exclusively on “big bang” food resources were sampled intermittently, with significant temporal variation in the capture rates of two species (Cynopterus horsfieldi and Megaerops ecaudatus). Significant variation in the capture rates of the remaining three species (Dyacopterus spadiceus, Eonycteris spelaea, and Rousettus amplexicaudatus) could not be detected due to small sample sizes. Since ephemeral “big bang” food resources were only sporadically available within the study area and were associated with large canopy trees and strangler figs, these results suggest that food abundance, or the availability of specific food items, may be important factors limiting local fruit bat species diversity in old‐growth Paleotropical rain forest. Thus, only three fruit bat species were locally resident within the forest throughout the study period. Therefore, further studies on the ranging behavior and habitat requirements of Malaysian fruit bats are required to assess the adequacy of existing reserves and protected areas. 相似文献
58.
Sharon L. Gursky 《International journal of primatology》1995,16(1):843-853
I present quantitative and qualitative data on infant caretaking behaviors collected during a preliminary field study of spectral
tarsiers (Tarsius spectrum),in a northern Sulawesi rain forest. The primary goal of the study is to identify the basic pattern of infant care in this
species. I studied tarsiers at Tangkoko-Dua Saudara Nature Reserve in Sulawesi, Indonesia, from May to July 1992. I observed
two infants, from two groups for a total of 96 hr using focal follows. During individual focal follows, ranging from 20 min
to 6 hr, I recorded behaviors at 5-min intervals. I also recorded distances of group members relative to the infant at 5-min
intervals. I subsampled the data at 35-min intervals to control for statistical autocorrelation between data points. Infants
were alone between 40 and 50% of the time. The two subadults were more frequently in proximity to the infant than the adult
males, the nonmatemal adult female, or the mothers were. This pattern of the subadults maintaining proximity to the infant
continued when the mothers were absent. These results suggest that subadults may be guarding or babysitting infants. It is
also possible that subadults are not traveling as far from the sleeping site as adults do and are therefore more likely to
be found in association with the infant. 相似文献
59.
60.
Selenoprotein P associates with endothelial cells in rat tissues 总被引:11,自引:0,他引:11
R. F. Burk Kristina E. Hill Martha E. Boeglin Ford F. Ebner Harold S. Chittum 《Histochemistry and cell biology》1997,108(1):11-15
Selenoprotein P is an extracellular heparin-binding protein that has been implicated in protecting the liver against oxidant
injury. Its location in liver, kidney, and brain was determined by conventional immunohistochemistry and confocal microscopy
using a polyclonal antiserum. Selenoprotein P is associated with endothelial cells in the liver and is more abundant in central
regions than in portal regions. It is also present in kidney glomeruli associated with capillary endothelial cells. Staining
of selenoprotein P in the brain is also confined to vascular endothelial cells. The heparin-binding properties of selenoprotein
P could be the basis for its binding to tissue. Its localization to the vicinity of endothelial cells is potentially relevant
to its oxidant defense function.
Accepted: 6 March 1997 相似文献