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Query: UNIPROT:P06889 (
Mol
)
630,302
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
In order to determine how T cell-presented peptides associate with the antigen binding sites (desetopes) of class I major histocompatibility complex (MHC) molecules and how they might be scavenged from an endogenous processing pathway for transfer to those molecules, we characterized the binding of two synthetic peptides restricted by HLA-B37 or HLA-A2 to class I MHC molecules and to cellular proteins of histotyped cell lines, by gel filtration and photo-affinity labeling techniques. In gel filtration binding studies, each peptide associated with immunopurified class I MHC molecules from cells with its restricting, histotype, but little was bound to class I MHC molecules from cells without the restricting histotype and none was bound to bovine
serum albumin
. After crosslinkage of a radioiodinated photoreactive derivative of influenza virus nucleoprotein peptide NP(336-355Y) and immunoprecipitations with antibodies to class I MHC molecules, that peptide was found to bind to immunopurified class I MHC molecules from HLA-B37+ but not HLA-B37- cells. Binding of the [125I]NP peptide increased from 6 to 12 hr of incubation and was competed by unlabeled, NP peptide but not by HLA-A2-restricted, influenza virus matrix MA(57-73). The principal microsomal membrane proteins binding [125I]NP were about 65, 45 and 33 kD.
Mol
Immunol
PMID:Binding of radioiodinated influenza virus peptides to class I MHC molecules and to other cellular proteins as analyzed by gel filtration and photoaffinity labeling. 206 16
An immunosorbent technique was developed to attenuate cross-reactivity of a polyclonal antiserum against a 4(2) (rho-carboxyphenylazo)-1,3,5[10]-estratrien-3,16 alpha,17 beta-triol-bovine
serum albumin
conjugate. The chromatographic separation of antiserum through stationary phases having either rho(carboxymethyl)phenylazo-phenol or rho(carboxymethyl)-phenylazo-2-naphthol side residues reduced the antiserum avidity, while increasing the apparent antiserum affinity and decreasing the residual cross-reactivities against heterologous ligands. The highly specific antiserum obtained allowed the development of a competitive binding assay over an extended analytical range, which opens up the possibility of direct measurement of estriol from the early pregnancy to delivery. The significance of the attenuation of antiserum cross-reactions after affinity chromatography is discussed with reference to epitope-paratope interaction in the case of small endogenous molecules like estrogens.
J Steroid Biochem
Mol
Biol 1991 Jul
PMID:Highly specific polyclonal antisera against estriol: cross-reactivity restriction following affinity chromatography. 206 67
A number of proteinases are induced and secreted into the culture medium of Tritirachium album Limber when the nitrogen source is limited to exogenous proteins. We have constructed a cDNA library using the polyadenylated RNA isolated during the nutritional induction with bovine
serum albumin
. A full-length clone of a gene for a new proteinase (named proteinase R) was identified from this library. This clone contained sequences coding for the 108-amino-acid prepro-leader as well as for the 279-amino-acid mature proteinase. Proteinase R apparently belongs to the subtilisin group of serine proteases that contains disulphide bonds. Homology between proteinase R and proteinase K was found to be about 87% at the nucleotide as well as at the amino acid level. The Brookhaven Protein Data Base co-ordinate file of proteinase K was used as a template to study the proteinase R substitutions in three-dimensional space. The majority of the substitutions of proteinase R with respect to proteinase K were found to be on the exterior of the protein model.
Mol
Microbiol 1990 Oct
PMID:Isolation and characterization of the gene encoding a novel, thermostable serine proteinase from the mould Tritirachium album Limber. 207 61
The development of cultured rabbit preimplantation embryos grown in standard media (Ham's F-10 or BSM II supplemented with bovine
serum albumin
(BSA) or homologous serum) or in Ham's medium supplemented with uterine flushings was compared. The uterine flushings derived from donors of 0.5-6 years of age. Uterine flushing supplemented media were used natively or after treatments like sterilization by filtration, lyophilization, three times freezing/thawing, heat denaturation, dialysis, or ultrafiltration. Compared with in vivo controls, embryonic growth was substantially reduced during in vitro culture, demonstrably by smaller diameters and impaired cell proliferation (measured by thymidine incorporation). The growth retardation was more pronounced in blastocysts (recovered at day 4 post coitum [p.c.]) than in morulae (recovered at day 3 p.c.). Development in uterine flushing media was notably better than in standard media but did not comply with in vivo development. Highest thymidine incorporation was observed in media with increased concentrations of uterine secretions and after sequential supplementation of flushings from subsequent progestational stages. Advanced donor ages, heating up to 80 degrees C, freezing, and lyophilizing did not affect incorporation data statistically significantly, whereas sterilization by filtration, ultrafiltration, and dialysis led to a significantly reduced thymidine incorporation in the cultured embryos. The positive effects of uterine flushing supplementation are attributed to the supply of components more adjusted to the needs of the cultured embryos and/or to a reduction of pathological effects in vitro like washing out of nutritive and regulatory components from the embryo into the surrounding culture medium.
Mol
Reprod Dev 1990 Nov
PMID:Development of preimplantation rabbit embryos in uterine flushing-supplemented culture media. 207 37
Binding equilibria of valproate (2-n-propyl-pentanoic acid anion) with defatted human
serum albumin
were studied by equilibrium dialysis in a 66 mM sodium phosphate buffer, pH 7.4, 37 degrees. Three hundred and fifty-six observed points for bound versus free valproate concentration were obtained and analyzed in terms of stepwise binding. It was found that the best fit resulted from a model in which 67% of the albumin was capable of binding valproate, whereas 33% did not bind. Thirty acceptable variants of the curve fitting were generated in order to assess the variation of the binding constants. The binding albumin component combines with three molecules of valproate with high affinity and with at least seven additional molecules that are loosely bound. Saturation of the protein cannot be reached. At very high concentrations of free valproate (above 10 mM) irreversible changes in the albumin take place, resulting in poor reproducibility in the amount of bound valproate. In the presence of palmitate, 0.5, 1, and 1.5 mol/mol of albumin, binding of valproate is decreased by a competitive mechanism. It is hypothesized that obesity, developing as a complication of valproate treatment of epilepsy, results from increased availability of long-chain fatty acids due to competitive valproate binding.
Mol
Pharmacol 1990 May
PMID:Valproate and palmitate binding to human serum albumin: an hypothesis on obesity. 211 Oct 5
In previous studies from this laboratory, a mediated transport system for long chain fatty acids was observed in rat renal basolateral membrane vesicles. Transport was measured in the absence of albumin and indicated the presence of a Na+ independent anion exchange mechanism. The present experiments were done to characterize renal transport of fatty acids derived from fatty acid-albumin complexes. 3H-palmitate uptake by brush border (BBMV) and basolateral membrane vesicles (BLMV) isolated from rat renal cortex was determined using a rapid filtration technique. All incubation media contained 100 microM 3H-palmitate complexed to 100 microM bovine
serum albumin
. Up to 65% of initially bound fatty acid-albumin complexes were displaceable by washing with solution containing 0.1% albumin. Total palmitate uptake was measured as the remaining non-displaceable radioactivity. In BBMV in low ionic strength (300 mM mannitol) or ionic buffers (100 mM mannitol + 100 mM NaCl or KCl), total palmitate uptake at 15 sec did not differ from equilibrium (60 min) values of 10-11 nmoles/mg protein. Uptake was primarily due to binding. A similar pattern was seen with BLMV in 300 mM mannitol buffer. In BLMV in 100 mM NaCl or KCl buffers, equilibrium uptake was 10-fold lower than at 15 sec. This suggests binding followed by cation-dependent translocation. If a putative FABPPM is involved in transport only, its presence should be confined to BLMV.
Mol
Cell Biochem
PMID:Renal palmitate transport: possible sites for interaction with a plasma membrane fatty acid-binding protein. 212 15
The N-glycan-processing inhibitors swainsonine (Sw) and deoxymannojirimycin (dMM) were used to study the influence of N-glycans on iodide organification in cultured porcine thyroid cells. Incubations with [125I]NaI were followed by determination of labeled trichloroacetic acid-insoluble material in culture media, follicular contents and cells. In controls, most of this material was in the follicular contents. With Sw and dMM, total acid-insoluble material was less than 10% of control. Iodide uptake was slightly inhibited and hydrogen peroxide release was not affected by inhibitors. Cell-surface thyroid peroxidase (TPO) activity, assayed by its ability to iodinate bovine
serum albumin
, was strongly inhibited. Pronase glycopeptide analysis indicated that with drugs the content in complex-type N-glycans was strongly decreased while that in hybrid or oligomannosidic type was increased. In conclusion, inhibition of N-glycan processing prevents iodide organification in cultured porcine thyroid cells by decreasing the recovery of cell-surface TPO activity.
Mol
Cell Endocrinol 1990 Oct 22
PMID:Inhibition of N-glycan processing affects iodide organification in porcine thyroid cells. 214 33
The bioactivity of an inhaled particulate is likely to be influenced significantly by the adsorption of proteins and lipids from the fluid lining the respiratory tract. We have shown previously in vitro that the presence of immunoglobulin G, a protein constituent of the lining fluid, significantly and specifically enhances the response of the guinea pig alveolar macrophage to chrysotile, but not crocidolite, asbestos. In order to understand the differential response of the guinea pig alveolar macrophage to chrysotile and crocidolite, as well as to obtain a better understanding of the factors that determine the adsorption of proteins onto asbestos, we have quantitated the adsorption of model proteins, including IgG, onto asbestos. The apparent surface density to which a given protein adsorbed was found to be influenced significantly by the surface charge of the asbestos, by the net protein charge, i.e., its isoelectric point (pI), and most dramatically, by the presence of other proteins. Thus, for example, even when guinea pig IgG (gpIgG; pI approximately 8.5) and bovine
serum albumin
(BSA; pI = 4.8) were present together at a high molar ratio of BSA to gpIgG, gpIgG was selectively adsorbed on negatively charged crocidolite asbestos. An analogous result was found for the adsorption of gpIgG and cytochrome c (CYT; pI = 10) on positively charged chrysotile asbestos, i.e., even at high molar ratios of CYT to gpIgG, gpIgG was selectively adsorbed. This latter result provides an explanation for the differential bioactivities of chrysotile and crocidolite asbestos previously noted in vitro.(ABSTRACT TRUNCATED AT 250 WORDS)
Am J Respir Cell
Mol
Biol 1990 May
PMID:Modification of asbestos bioactivity for the alveolar macrophage by selective protein adsorption. 216 Feb 55
Oxidant-induced injury is associated with breakdown of interstitial collagen and the accumulation of inflammatory cells in the lungs. In previous studies, we demonstrated that phagocyte accumulation is mediated, in part, by chemotactic factors generated from damaged collagen. To determine if alveolar macrophages also mediate the migration of polymorphonuclear leukocytes (PMN) into the lungs, we examined the release of chemotactic factors from alveolar macrophages treated with native or synthetic collagenous peptides. These included fragments of bovine collagen digested with bacterial collagenase (CG) or cyanogen bromide (CB) as well as small molecular weight synthetic polypeptides containing proline (Pro), glycine (Gly), and hydroxyproline (Hyp), the major amino acids that comprise collagen. We found a dose- and time-related generation of PMN chemotactic activity by collagen peptide-treated macrophages. The maximum activity was released 72 h after pretreatment of macrophages for 1 to 3 h with 0.1 to 1 microM CG-, CB-, or (Pro-Pro-Gly)5-peptides. The native peptides derived from CG-digested collagen were more active than synthetic peptides containing Pro and Gly. Neither trypsin digests of bovine
serum albumin
nor synthetic peptides containing Hyp stimulated chemotactic factor release from macrophages. The alveolar macrophage-derived chemotactic factor was found to lose activity when dialyzed and after heat or trypsin treatment. The release of PMN-activating factors by collagen peptide-treated macrophages was also examined. Alveolar macrophage-conditioned medium was found to stimulate PMN production of reactive oxygen intermediates as well as elastase and gelatinase.(ABSTRACT TRUNCATED AT 250 WORDS)
Am J Respir Cell
Mol
Biol 1990 May
PMID:Activation of neutrophils by factors released from alveolar macrophages stimulated with collagen-like polypeptides. 216 Feb 56
Side-chain cleavage (SCC) of endogenous cholesterol in adrenal mitochondria isolated from ACTH-treated rats indicates that the size of the reactive cholesterol pool depends on the reducing precursor. At optimal concentrations of reductant, this pool was typically at least 2 times greater for isocitrate than for succinate. Succinate-supported reactions were rapidly completed, were highly sensitive to a 2-min preincubation, and failed to deplete spectrally detected P-450SCC-cholesterol complexes. Cholesterol SCC with 1 mM isocitrate exhibited 2-3 times more fast-phase metabolism, a pronounced slow phase, insensitivity to preincubation, and 60% depletion of spectrally detected cholesterol-P-450SCC complexes. Addition of bovine
serum albumin
(BSA) and EDTA, either during homogenization or directly to the incubation, prevented preincubation losses in response to succinate and removed most of the difference between succinate and isocitrate activities. This effect of BSA/EDTA was reversed within 5 min by octanoate by a mechanism that was enhanced by Ca2+. These distinct reductant characteristics suggest that only a subpopulation of mitochondria or of pools of activity within individual mitochondria can support cholesterol SCC with succinate while isocitrate is necessary for the remainder. The rapid responses of succinate-supported metabolism to preincubation or to octanoate suggest depletion of a critical factor for cholesterol metabolism. Metabolism of added 20 alpha-hydroxycholesterol or deoxycorticosterone established that NADPH remained fully available after succinate-supported cholesterol metabolism had stopped or after preincubation. Cessation of pregnenolone formation, therefore, results from a failure to supply cholesterol, not inadequate NADPH. The preincubation effect suggests loss of an energy-dependent component that enhances this supply of cholesterol. One possibility tested was that GTP, an activator of intermembrane cholesterol transfer (Xu et al. (1989) J. Biol. Chem. 264, 17674-17680), was being lost. Added GTP slightly activated succinate-supported pregnenolone production but did not prevent preincubation-induced losses. alpha-Ketoglutarate, which can generate matrix GTP, is an effective reductant that, in combination with succinate, prevents preincubation-induced losses.
Mol
Cell Endocrinol 1990 Oct 22
PMID:Heterogeneous pools of cholesterol side-chain cleavage activity in adrenal mitochondria from ACTH-treated rats: differential responses to different reducing precursors. 217 27
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