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81.
The catalytic domains of murine Golgi alpha1,2-mannosidases IA and IB that
are involved in N-glycan processing were expressed as secreted proteins in
P.pastoris . Recombinant mannosidases IA and IB both required divalent
cations for activity, were inhibited by deoxymannojirimycin and
kifunensine, and exhibited similar catalytic constants using
Manalpha1,2Manalpha-O-CH3as substrate. Mannosidase IA was purified as a 50
kDa catalytically active soluble fragment and shown to be an inverting
glycosidase. Recombinant mannosidases IA and IB were used to cleave
Man9GlcNAc and the isomers produced were identified by high performance
liquid chromatography and proton-nuclear magnetic resonance spectroscopy.
Man9GlcNAc was rapidly cleaved by both enzymes to Man6GlcNAc, followed by a
much slower conversion to Man5GlcNAc. The same isomers of Man7GlcNAc and
Man6GlcNAc were produced by both enzymes but different isomers of
Man8GlcNAc were formed. When Man8GlcNAc (Man8B isomer) was used as
substrate, rapid conversion to Man5GlcNAc was observed, and the same
oligosaccharide isomer intermediates were formed by both enzymes. These
results combined with proton-nuclear magnetic resonance spectroscopy data
demonstrate that it is the terminal alpha1, 2-mannose residue missing in
the Man8B isomer that is cleaved from Man9GlcNAc at a much slower rate.
When rat liver endoplasmic reticulum membrane extracts were incubated with
Man9GlcNAc2, Man8GlcNAc2was the major product and Man8B was the major
isomer. In contrast, rat liver Golgi membranes rapidly cleaved
Man9GlcNAc2to Man6GlcNAc2and more slowly to Man5GlcNAc2. In this case all
three isomers of Man8GlcNAc2were formed as intermediates, but a distinctive
isomer, Man8A, was predominant. Antiserum to recombinant mannosidase IA
immunoprecipitated an enzyme from Golgi extracts with the same specificity
as recombinant mannosidase IA. These immunodepleted membranes were enriched
in a Man9GlcNAc2to Man8GlcNAc2- cleaving activity forming predominantly the
Man8B isomer. These results suggest that mannosidases IA and IB in Golgi
membranes prefer the Man8B isomer generated by a complementary mannosidase
that removes a single mannose from Man9GlcNAc2.
相似文献
82.
Crystallization and preliminary x-ray investigation of purine-nucleoside phosphorylase from Escherichia coli 总被引:2,自引:0,他引:2
W J Cook S E Ealick T A Krenitsky J D Stoeckler J R Helliwell C E Bugg 《The Journal of biological chemistry》1985,260(24):12968-12969
Crystals of purine-nucleoside phosphorylase from Escherichia coli have been grown from solutions of ammonium sulfate. The crystals are hexagonal with space group P6(1)22 or P6(5)22; the axes are alpha = 106.5 A and c = 241.3 A. The crystals are moderately stable to x-rays and diffract beyond 3.0-A resolution. It appears that the molecule, which is a hexamer, utilizes the 2-fold symmetry of the space group, resulting in three subunits/asymmetric unit. 相似文献
83.
The involvement of gibberellin 20-oxidase genes in phytochrome-regulated petiole elongation of Arabidopsis 下载免费PDF全文
Long day (LD) exposure of rosette plants causes rapid stem/petiole elongation, a more vertical growth habit, and flowering; all changes are suggestive of a role for the gibberellin (GA) plant growth regulators. For Arabidopsis (Arabidopsis thaliana) L. (Heynh), we show that enhancement of petiole elongation by a far-red (FR)-rich LD is mimicked by a brief (10 min) end-of-day (EOD) FR exposure in short day (SD). The EOD response shows red (R)/FR photoreversibility and is not affected in a phytochrome (PHY) A mutant so it is mediated by PHYB and related PHYs. FR photoconversion of PHYB to an inactive form activates a signaling pathway, leading to increased GA biosynthesis. Of 10 GA biosynthetic genes, expression of the 20-oxidase, AtGA20ox2, responded most to FR (up to a 40-fold increase within 3 h). AtGA20ox1 also responded but to a lesser extent. Stimulation of petiole elongation by EOD FR is reduced in a transgenic AtGA20ox2 hairpin gene silencing line. By contrast, it was only in SD that a T-DNA insertional mutant of AtGA20ox1 (ga5-3) showed reduced response. Circadian entrainment to a daytime pattern provides an explanation for the SD expression of AtGA20ox1. Conversely, the strong EOD/LD FR responses of AtGA20ox2 may reflect its independence of circadian regulation. While FR acting via PHYB increases expression of AtGA20ox2, other GA biosynthetic genes are known to respond to R rather than FR light and/or to other PHYs. Thus, there must be different signal transduction pathways, one at least showing a positive response to active PHYB and another showing a negative response. 相似文献
84.
J Yariv A J Kalb J R Helliwell M Z Papiz E R Bauminger I Nowik 《The Journal of biological chemistry》1988,263(27):13508-13510
We report here the preparation of iron-containing concanavalin A. It has a protein-to-iron ratio of 2.0, and the iron compound it contains is particulate with an average diameter of 85 A. Iron-containing concanavalin A interacts reversibly with dextran and with methyl alpha-D-glucoside. The molecular basis of these findings is discussed and a possible mechanism suggested where one of the molecular forms of concanavalin A has the structure of an apoferritin into which iron is deposited in the form of ferrihydrite. 相似文献
85.
D Lederoun P Vandewalle AA Brahim J Moreau PA Lalèyè 《African Journal of Aquatic Science》2016,41(2):151-160
Growth, mortality, recruitment and relative yield per recruit of Sarotherodon galilaeus galilaeus from Lakes Doukon and Togbadji were studied. Data on total length, total weight and sex were recorded on a monthly basis between January and December 2013 for S. g. galilaeus captured by local fishers. The estimated asymptotic lengths L∞ were 26.2 and 23.6?cm for Lakes Doukon and Togbadji, respectively, while the growth rate K was 0.73 in Lake Doukon and 0.87 in Lake Togbadji. Estimates of fishing mortality, 0.27 and 0.47 y?1 for Doukon and Togbadji, respectively, were low relative to natural mortality, 1.51 and 1.74 y?1, respectively. Sizes at first sexual maturity were 12.8 and 13.2?cm for females and males, respectively, in Lake Doukon, and 11.5 and 12.4?cm for females and males, respectively, in Lake Togbadji. The size at first capture was estimated at 13.3 and 12.7?cm for Lakes Doukon and Togbadji, respectively, which, in the light of the size at maturity estimates, indicates that fish spawn at least once before capture. The current exploitation rates of 0.15 for Lake Doukon and 0.21 for Lake Togbadji suggest that their stocks of S. g. galilaeus are not overexploited in either lake. 相似文献
86.
87.
RC Pritchett AM Al-Nawaiseh KK Pritchett V Nethery PA Bishop JM Green 《Biology of sport / Institute of Sport》2015,32(3):249-254
Sweat production is crucial for thermoregulation. However, sweating can be problematic for individuals with spinal cord injuries (SCI), as they display a blunting of sudomotor and vasomotor responses below the level of the injury. Sweat gland density and eccrine gland metabolism in SCI are not well understood. Consequently, this study examined sweat lactate (S-LA) (reflective of sweat gland metabolism), active sweat gland density (SGD), and sweat output per gland (S/G) in 7 SCI athletes and 8 able-bodied (AB) controls matched for arm ergometry VO2peak. A sweat collection device was positioned on the upper scapular and medial calf of each subject just prior to the beginning of the trial, with iodine sweat gland density patches positioned on the upper scapular and medial calf. Participants were tested on a ramp protocol (7 min per stage, 20 W increase per stage) in a common exercise environment (21±1°C, 45-65% relative humidity). An independent t-test revealed lower (p<0.05) SGD (upper scapular) for SCI (22.3 ±14.8 glands · cm−2) vs. AB. (41.0 ± 8.1 glands · cm−2). However, there was no significant difference for S/G between groups. S-LA was significantly greater (p<0.05) during the second exercise stage for SCI (11.5±10.9 mmol · l−1) vs. AB (26.8±11.07 mmol · l−1). These findings suggest that SCI athletes had less active sweat glands compared to the AB group, but the sweat response was similar (SLA, S/G) between AB and SCI athletes. The results suggest similar interglandular metabolic activity irrespective of overall sweat rate. 相似文献
88.
Joanna Maleszka Andrew B. Barron Paul G. Helliwell Ryszard Maleszka 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》2009,195(8):733-740
We examined the effects of behaviour, age and social environment on mushroom body volume in adult bees. The mushroom bodies
are regions of the central brain important for sensory integration and learning. Their volume was influenced by behaviour
throughout life: always larger in forager bees than age-matched nurse bees, even in old bees up to 93 days of age as adults.
Mushroom body development was influenced by the social environment in the first 8 days of adult life, with different environments
having markedly different effects on mushroom body size. Compared to hive-reared bees, isolation slowed mushroom body growth,
but bees reared in isolation confined with a single dead bee showed a dramatic increase in mushroom body volume comparable
to that seen in active foragers. Despite their precocious mushroom body development, these bees did not show improved performance
in an olfactory learning test. Since simple environmental manipulations can both accelerate and delay mushroom body growth
in young bees, and since mushroom body volume is sensitive to behaviour throughout life, the honey bee has great potential
as a model for exploring the interactions between environment, behaviour and brain structure. 相似文献
89.
90.