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101.
Although considerable information is available on the oligomeric states of the AE1 (band 3) anion exchanger, little is known about the physiological state of the polypeptides encoded by the nonerythroid AE genes, AE2 and AE3. We have previously characterized the proteolytic susceptibility of native pig gastric AE2. In the course of studies in which pig gastric membranes were treated with the AE2 transport antagonist, DIDS, we noted evidence for cross-linking of AE2 proteolytic fragments to higher-order oligomeric forms. We have characterized the ability of DIDS and of selected N-hydroxysuccinimide cross-linking agents to increase the proportion of SDS-resistant oligomers of pig gastric AE2 and its proteolytic fragments. Cross-linking exhibited time and concentration dependence. N-Terminal protein sequencing proved that DIDS treatment created AE2 homodimers. Putative homotetramers were also observed. Protomers were cross-linked via residues within the C-terminal 40 kDa of AE2. Prior proteolytic cleavage of AE2 in membranes resulted in decreased yield of subsequently cross-linked products. AE2 cross-linking could not be detected in membranes pretreated by hypotonic wash and freeze-thaw. The results are interpreted in light of the deduced amino acid sequence of the transmembrane domain of pig AE2. 相似文献
102.
Elena V. Navolotskaya Yulia A. Kovalitskaya Vladimir B. Sadovnikov Yury A. Zolotarev Alexander A. Kolobov Vladimir V. Yurovsky Valery M. Lipkin 《International journal of peptide research and therapeutics》2008,14(1):10-15
Synthetic adrenocorticotropic hormone (ACTH)-like octapeptide leukocorticotropin (GKVLKKRR), corresponding to the amino acid
sequence 81–88 of pro-interleukin-1α, was labeled with tritium (specific activity of 22 Ci/mmol) and was found to bind to
rat adrenal cortex membranes with high affinity and specificity (K
d = 2.2 ± 0.1 nM). Synthetic 125I-labeled ACTH fragment 11–24 was also obtained (specific activity of 98 Ci/mmol) and shown to bind to ACTH receptor on rat
adrenal cortex with high affinity (K
d = 1.8 ± 0.1 nM). Unlabeled leukocorticotropin was found to actively replace 125I-labeled ACTH (11–24) in the receptor-ligand complex (K
i = 2.0 ± 0.1 nM). Leukocorticotropin at concentration range of 1–1000 nМ did not affect the adenylate cyclase activity in
adrenocortical membranes. Thus, leukocorticotropin is an antagonist of ACTH receptor.
ACTH-like peptide GKVLKKRR is an antagonist of ACTH
This work financeed by the Russian Foundation for Basic Research (grant No. 05-04-48060), by the programs Leading Scientific
Schools (grant No. 312.2003.4), Molecular and Cellular Biology (chairman V.M. Lipkin), and Naukogrady (grant No. 04-04-97200),
and by the International Science and Technology Center (project No. 2615). V.V. Yurovsky is supported by the American Heart
Association (grant No. 0555415U). 相似文献
103.
Yu. A. Kovalitskaya A. A. Kolobov E. A. Kampe-Nemm Yu. A. Zolotarev V. V. Yurovskii V. B. Sadovnikov V. M. Lipkin E. V. Navolotskaya 《Russian Journal of Bioorganic Chemistry》2008,34(1):24-29
Tritium-labeled synthetic fragments of human adrenocorticotropic hormone (ACTH) [3H]ACTH (11–24) and [3H]ACTH (15–18) with a specific activity of 22 and 26 Ci/mmol, respectively, were obtained. It was found that [3H]ACTH-(11–24) binds to membranes of the rat adrenal cortex with high affinity and high specificity (K d 1.8 ± 0.1 nM). Twenty nine fragments of ACTH (11–24) were synthesized, and their ability to inhibit the specific binding of [3H]ACTH (11–24) to adrenocortical membranes was investigated. The shortest active peptide was found to be an ACTH fragment (15–18) (KKRR) (K i 2.3 ± 0.2 nM), whose [3H] labeled derivative binds to rat adrenocortical membranes (K d 2.1 ± 0.1 nM) with a high affinity. The specific binding of [3H]ACTH-(15–18) was inhibited by 100% by unlabeled ACTH (11–24) (K i 2.0 ± 0.1 nM). ACTH (15–18) in the concentration range of 1–1000 nM did not affect the adenylate cyclase activity of adrenocortical membranes and, therefore, is an antagonist of the ACTH receptor. 相似文献
104.
I. P. Ashmarin K. E. Baglikova S. E. Edeeva Yu. A. Zolotarev V. S. Kozik A. K. Dadayan E. M. Dorokhova L. Yu. Alfeeva L. A. Andreeva G. N. Kopylova T. S. Pavlov B. V. Vaskovsky V. K. Meshavkin O. Yu. Sokolov N. V. Kost A. A. Zozulya G. E. Samonina N. F. Myasoedov 《Russian Journal of Bioorganic Chemistry》2008,34(4):415-420
The distribution of the glyprolines, Pro-Gly-Pro and Thr-Lys-Pro-Arg-Pro-Gly-Pro (Selanc), was analyzed and compared in tissues of rat organs after different ways of their administration using the peptides uniformly labeled with tritium. Comparative data on changes of concentrations of the peptides in the rat organs after their intraperitoneal, intranasal, intragastric, and intravenous administration are given. The intranasal administration of both peptides was shown to be optimal for delivery of glyprolines molecules in the CNS. A high affinity of the studied glyprolines for gastric tissues was found for all the ways of their administration. We suggest that high efficacy of action of glyprolines on homeostasis of the gastric mucosa was partially provided by accumulation of these peptides (to high concentrations) in gastric tissues. 相似文献
105.
106.
Kolobov AA Kolodkin NI Zolotarev IuA Tathill C Navolotskaia EV 《Bioorganicheskaia khimiia》2008,34(1):43-49
The tritium-labeled dipeptide bestim (gamma-D-Glu-L-Trp) with a specific activity of 45 Ci/mmol was obtained by high-temperature solid-state catalytic isotope exchange. It was found that [3H]bestim binds with a high affinity to murine peritoneal macrophages (Kd 2.1 +/- 0.1 nM) and thymocytes (Kd 3.1 +/- 0.2 nM), as well as with plasma membranes isolated from these cells (Kd 18.6 +/- 0.2 and 16.7 +/- 0.3 nM, respectively). The specific binding of [3H]bestim to macrophages and thymocytes was inhibited by the unlabeled dipeptide thymogen (L-Glu-L-Trp) (Ki 0.9 +/- 0.1 and 1.1 +/- 0.1 nM, respectively). After treatment with trypsin, macrophages and thymocytes lost the ability to bind [3H]bestim. Bestim in the concentration range of 10(-10) to 10(-6) M reduced the adenylate cyclase activity in the membranes of murine macrophages and thymocytes. 相似文献
107.
Many papers dealing with nonindigenous species in the Black Sea do not mention the polychaete Polydora limicola Annenkova, a new invader in the Pontic-Azov Basin. In this connection, we will analyze here the taxonomic status of this species and discuss the history of its introduction and distribution in the northwestern Black Sea and related coastal lagoons. It is shown that P. limicola is now one of the most common species of the benthos in the Black Sea, especially in estuarine areas and coastal lagoons. The naturalization of this species resulted in changes in the structure of some bottom biocenoses of this region. 相似文献
108.
E. V. Navolotskaya V. B. Sadovnikov D. V. Zinchenko V. I. Vladimirov Y. A. Zolotarev 《Russian Journal of Bioorganic Chemistry》2017,43(6):673-677
The 125I-labeled B-subunit of the cholera toxin ([125I]CT-B, specific activity of 98 Ci/mmol) was prepared. This subunit was shown to be bound to the membranes which were isolated from epithelial cells of a mucous tunic of the rat thin intestine with high affinity (K d = 3.7 nM). The binding of the labeled protein was inhibited by the unlabeled α2-interferon (IFN-α2), α1-thymosin, (TM-α1), and the LKEKK synthetic peptide corresponding to the 16–20 sequence of TM-α1 and the 131–135 sequence of human IFN-α2 (Ki 1.0, 1.5, and 2.0 nM, respectively), whereas the KKEKL unlabeled synthetic peptide did not inhibit the binding (K i > 100 μМ). The LKEKK peptide and CT-B were shown to dose-dependently increase an activity of the soluble guanylate cyclase (sGC) in the concentration range from 10 to 1000 nM. Thus, the binding of TM- α1, IFN-α2, and the LKEKK peptide to the CT-B receptor on a surface of the epithelial cells of the mucous tunic of the rat thin intestine resulted in an increase in the intracellular level of cGMP. 相似文献
109.
N. V. Kost O. Yu. Sokolov M. V. Gabaeva Yu. A. Zolotarev V. V. Malinin V. Kh. Khavinson 《Biology Bulletin》2003,30(4):351-353
The effects of new peptide bioregulators—Livagen (Lys-Glu-Asp-Ala) and Epitalon (Ala-Glu-Asp-Gly)—on the endogenous opioid system was studied. In particular, attention was focused on their ability to change the activity of enkephalin-degrading enzymes of blood serum and interact with opioid receptors of the brain membrane fraction. Enkephalinase activity was assayed in vitro by the rate of 3H-Leu-enkephalin hydrolysis in the presence of Livagen and Epitalon. These peptides inhibited enkephalin-degrading enzymes of human serum. Livagen proved to be more efficient than some well-known peptidase inhibitors, such as puromycin, leupeptin, and D-PAM. The dose–inhibitory effect curves for Livagen and Epitalon were plotted; their IC50 equaled 20 and 500 M, respectively. The interaction between the peptides and opioid receptors was estimated using a radioreceptor method with [3H][D-Ala2, D-Leu5]-enkephalin. No interaction was observed between the peptides and - or -opioid receptors of the membrane fraction from the rat brain. 相似文献
110.
Yulia N. Nekrasova Yury A. Zolotarev Elena V. Navolotskaya 《Journal of peptide science》2012,18(2):83-87
Two selective agonists of nonopioid β‐endorphin receptor, synthetic peptides TPLVTLFK (octarphin) and SLTCLVKGFY (immunorphin), were labeled with tritium to specific activity of 29 and 25 Ci/mmol, respectively. Both labeled peptides were found to bind to high‐affinity naloxone‐insensitive binding sites on the membranes isolated from the rat myocardium (Kd = 2.0 ± 0.2 and 2.5 ± 0.3 nM, respectively). The [3H]octarphin specific binding to the myocardial membranes was inhibited by unlabeled β‐endorphin (Ki = 1.9 ± 0.2 nM) and immunorphin (Ki = 2.2 ± 0.3 nM). The [3H]immunorphin specific binding with the membranes was inhibited by unlabeled β‐endorphin (Ki = 2.3 ± 0.3 nM) and octarphin (Ki = 2.4 ± 0.3 nM). The binding specificity study revealed that these binding sites were insensitive not only to naloxone but also to α‐endorphin, γ‐endorphin, [Met5]enkephalin and [Leu5]enkephalin. Thus, β‐endorphin, immunorphin and octarphin bind to the common high‐affinity naloxone‐insensitive receptor of the rat myocardial membranes. Copyright © 2011 European Peptide Society and John Wiley & Sons, Ltd. 相似文献