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11.
Elizabeth M. Harris Shirley C. Tucker Lowell E. Urbatsch 《American journal of botany》1991,78(1):108-121
The order of floral initiation and subsequent organogeny of Erigeron philadelphicus L. (Asteraceae: Astereae) was found to deviate from the acropetal pattern generally reported for the Asteraceae. Light micrographs show periclinal divisions in the first, second, and deeper subsurface layers of cells on the flanks of the inflorescence apex as the earliest evidence of floral initiation. Scanning electron microscope micrographs indicate that the disk flowers appear first and arise as small protuberances approximately one-third of the way up the previously and undifferentiated highly convex inflorescence apex. A succession of disk flowers arises acropetally in a complex anthotaxy characterized by about 21 dextrorse and 12–15 sinistrorse parastichies (although this pattern is obscured at the apex). After one to three disk flowers have been initiated in each parastichy, the first ray flower initials can be seen to initiate in sites proximal to the oldest and largest disk flowers. Additional ray flowers then initiate basipetally following the dextrorse parastichies established by the disk flowers. Overall floral initiation on the inflorescence apex proceeds acropetally for the disk flowers and basipetally for the ray flowers until the available space is filled. Floral development adheres to the same plan—proceeding bidirectionally on the inflorescence meristem with the oldest and most complete flowers of both types located on the equator established at initiation. 相似文献
12.
Chloupková M Pickert A Lee JY Souza S Trinh YT Connelly SM Dumont ME Dean M Urbatsch IL 《Biochemistry》2007,46(27):7992-8003
Human ATP-binding cassette (ABC) transporters comprise a family of 48 membrane-spanning transport proteins, many of which are associated with genetic diseases or multidrug resistance of cancers. In this study, we present a comprehensive approach for the cloning, expression, and purification of human ABC transporters in the yeast Pichia pastoris. We analyzed the expression of 25 proteins and demonstrate that 11 transporters, including ABCC3, ABCB6, ABCD1, ABCG1, ABCG4, ABCG5, ABCG8, ABCE1, ABCF1, ABCF2, and ABCF3, were expressed at high levels comparable to that of ABCB1 (P-glycoprotein). As an example of the purification strategy via tandem affinity chromatography, we purified ABCC3 (MRP3) whose role in the transport of anticancer drugs, bile acids, and glucuronides has been controversial. The yield of ABCC3 was 3.5 mg/100 g of cells in six independent purifications. Purified ABCC3, activated with PC lipids, exhibited significant ATPase activity with a Vmax of 82 +/- 32 nmol min-1 mg-1. The ATPase activity was stimulated by bile acids and glucuronide conjugates, reaching 170 +/- 28 nmol min-1 mg-1, but was not stimulated by a variety of anticancer drugs. The glucuronide conjugates ethinylestradiol-3-glucuronide and 17beta-estradiol-17-glucuronide stimulated the ATPase with relatively high affinities (apparent Km values of 2 and 3 microM, respectively) in contrast to bile acids (apparent Km values of >130 microM), suggesting that glucuronides are the preferred substrates for this transporter. Overall, the availability of a purification system for the production of large quantities of active transporters presents a major step not only toward understanding the role of ABCC3 but also toward future structure-function analysis of other human ABC transporters. 相似文献
13.
Kang Yun Hee Ji Na Young Lee Chung Il Lee Hee Gu Kim Jae Wha Yeom Young IL Kim Dae Ghon Yoon Seung Kew Kim Jong Wan Park Pil Je Song Eun Young 《Amino acids》2011,40(3):1003-1013
Amino Acids - Endothelial cell-specific molecule-1 (ESM-1) is a secretory proteoglycan comprising a mature polypeptide of 165 amino acids and a single dermatan sulfate. The aim of this study was to... 相似文献
14.
Nucleotide-induced structural changes in P-glycoprotein observed by electron microscopy 总被引:3,自引:0,他引:3
P-glycoprotein (Pgp) is an ATP hydrolysis driven multidrug efflux pump, which, when overexpressed in the plasma membrane of certain cancers, can lead to the failure of chemotherapy. Previously, we have presented a projection structure of nucleotide-free mouse Pgp from electron microscopic images of lipid monolayer-generated two-dimensional crystals ( Lee, J. Y., Urbatsch, I. L., Senior, A. E., and Wilkens, S. (2002) J. Biol. Chem. 277, 40125-40131 ). Here we have analyzed the structure of cysteine-free human Pgp from two-dimensional crystals that were generated with the same lipid-monolayer technique in the absence and presence of various nucleotides. The images show that human Pgp has a similar structure to the mouse protein. Furthermore, the analysis of projection structures obtained under different nucleotide conditions suggests that Pgp can exist in at least two major conformations, one of which shows a central cavity between the N- and C-terminal halves of the molecule and another in which the two halves have moved sideways, thereby closing the central cavity. Intermediate conformations were observed for some nucleotide/vanadate combinations. A low-resolution, three-dimensional model of human Pgp was calculated from tilted specimen crystallized in the presence of the non-hydrolyzable nucleotide analog, adenosine 5'-O-(thiotriphosphate). The structural analysis presented here adds to the emerging picture that multidrug ABC transporters function by switching between two major conformations in a nucleotide-dependent manner. 相似文献
15.
Alfonso G. Ober Leovigildo Quijano Lowell E. Urbatsch Nikolaus H. Fischer 《Phytochemistry》1984,23(6):1289-1292
Chemical analysis of Calea subcordata yielded three new guaianolides, 8-epi-isobutyrylrupicolin A, 8-epi-isobutyrylrupicolin B and subcordatolide A, which represent the first guaianolide-type sesquiterpene lactones reported in the genus Calea. The structures of the new compounds were established by chemical and spectroscopic methods. 相似文献
16.
17.
Schölz C Parcej D Ejsing CS Robenek H Urbatsch IL Tampé R 《The Journal of biological chemistry》2011,286(15):13346-13356
The transporter associated with antigen processing (TAP) plays a key role in adaptive immunity by translocating proteasomal degradation products from the cytosol into the endoplasmic reticulum lumen for subsequent loading onto major histocompatibility (MHC) class I molecules. For functional and structural analysis of this ATP-binding cassette complex, we established the overexpression of TAP in the methylotrophic yeast Pichia pastoris. Screening of optimal solubilization and purification conditions allowed the isolation of the heterodimeric transport complex, yielding 30 mg of TAP/liter of culture. Detailed analysis of TAP function in the membrane, solubilized, purified, and reconstituted states revealed a direct influence of the native lipid environment on activity. TAP-associated phospholipids, essential for function, were profiled by liquid chromatography Fourier transform mass spectrometry. The antigen translocation activity is stimulated by phosphatidylinositol and -ethanolamine, whereas cholesterol has a negative effect on TAP activity. 相似文献
18.
19.
Alan E. Senior Marwan K. Al-Shawi Ina L. Urbatsch 《Journal of bioenergetics and biomembranes》1995,27(1):31-36
ATPase activity of multidrug-resistance protein (P-glycoprotein, Pgp) from Chinese hamster ovary cells was studied. Catalytic characteristics were established for Pgp both in its natural plasma membrane environment and in purified reconstituted protein. Generally the two preparations of Pgp behaved similarly, and demonstrated low affinity for MgATP, low nucleotide specificity, preference for Mg-nucleotide, and pH optimum near 7.5. A high-affinity binding site involved in catalysis was not apparent. Effective covalent inactivators were NBD-C1, NEM, 8-azido-ATP, and 2-azido-ATP. DCCD, FITC, and pyridoxal phosphate were only weakly inhibitory. Lipid composition was found to affect the degree of drug stimulation of ATPase in purified reconstituted Pgp, suggesting that the lipid environment affects coupling between drug-binding and catalytic sites, and that Pgp expressed in different tissues could show different functional characteristics. 相似文献
20.
The isolation and structure elucidation of two new guaianolides, solidaginolides A(1) and B(5), from Calea solidaginea are reported. The structures of the new compounds were established by spectroscopic methods. 相似文献