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The pH and salt dependences of the interaction of phosphatidic acid, phosphatidylserine, and stearic acid with myelin proteolipid apoprotein (PLP) in dimyristoylphosphatidylcholine (DMPC) recombinants have been studied by electron spin resonance spectroscopy, using spin-labeled lipids. The two-component spin-label spectra have been analyzed both by spectral subtraction and by simulation using the exchange-coupled Bloch equations to give the fraction of lipids motionally restricted by the protein and the rate of lipid exchange between the fluid and motionally restricted lipid populations. For stearic acid, phosphatidic acid, and phosphatidylserine, the fraction of motionally restricted spin-label increases with increasing pH, with pKa's of 7.7, 7.6, and ca. 9.4, respectively. The corresponding pKa's for the bulk lipid regions of the bilayer are estimated, from changes in the ESR spectra, to be 6.7, 7.4, and 11, respectively. In the dissociated state at pH 9.0, the fraction of motionally restricted component decreases with increasing salt concentration, reaching an approximately constant value at [NaCl] = 0.5-1.0 M for all three negatively charged lipids. The net decreases for stearic acid and phosphatidic acid are considerably smaller (by ca. 30%) than those obtained on protonating the two lipids, whereas for phosphatidylserine the fraction of motionally restricted lipid in high salt is reduced to that corresponding to phosphatidylcholine. For a fixed lipid/protein ratio, the on-rate for exchange at the lipid-protein interface is independent of the degree of selectivity and has a shallow temperature dependence, as expected for a diffusion-controlled process.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
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Aileen F. G. Bongat Rina Saksena Roberto Adamo Yukari Fujimoto Zenyu Shiokawa Dwight C. Peterson Koichi Fukase Willie F. Vann Pavol Kováč 《Glycoconjugate journal》2010,27(1):69-77
Using recombinant tetanus toxin HC fragment (rTT-HC) as carrier, we prepared multimeric bivalent immunogens featuring the synthetic hexasaccharide fragment of O-PS of Vibrio cholerae O:1, serotype Ogawa, in combination with either the synthetic hexasaccharide fragment of O-PS of Vibrio cholerae O:1, serotype Inaba, or a synthetic disaccharide tetrapeptide peptidoglycan fragment as adjuvant. The conjugation reaction
was effected by squaric acid chemistry and monitored in virtually real time by SELDI-TOF MS. In this way, we could prepare
well-defined immunogens with predictable carbohydrate–carrier ratio, whose molecular mass and the amount of each saccharide attached could be independently determined. The ability to prepare such neoglycoconjugates opens unprecedented possibilities
for preparation of conjugate vaccines for bacterial diseases from synthetic carbohydrates. 相似文献
189.
Monoclonal antibodies were produced to electrophoretically pure hydrogenase fromThiocapsa roseopersicina. Protein immunoelectroblotting was used to identify the hydrogenase-specific antibodies. Among the 18 monoclonal antibodies selected by enzyme immunoassay, three were found to react with highly immunogenic trace contaminating proteins. One cell line produced antibody that inhibitied hydrogenase activity. This was the first specific inhibitor of the hydrogenase function. The results suggest that monoclonal antibodies could provide valuable new informations about the enzyme structure as well. 相似文献
190.
Eduardo León-Ruiz Purificación Alcázar Eugenio Domínguez-Vilches Carmen Galán 《Aerobiologia》2011,27(1):37-50
The present study sought to determine which of the common Poaceae species in the study area contribute most to the Poaceae
pollen season curve, and to determine the phenological behaviour of the species studied. The different floral phenophases
in thirty-three Poaceae species common in and around the city of Córdoba (SW Iberian Peninsula) were checked periodically
over the period 2004–2006. Results showed that longer phenological ranges were recorded in the coolest and wettest year, and
shorter ranges in the warmest and driest year. Moreover, ranges varied as a function of altitude: populations in lower-lying
areas flowered earlier than those at higher altitudes. The results, taken in conjunction with the findings of preliminary
research into potential pollen production, showed that probably only four of the Poaceae species studied—Dactylis glomerata, Lolium rigidum, Trisetaria panicea and Vulpia geniculata—were major contributors to the Poaceae airborne pollen curve. 相似文献