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排序方式: 共有304条查询结果,搜索用时 15 毫秒
1.
George Theodoropoulos Coralie A. Carothers Carraway Kermit L. Carraway 《Journal of cellular biochemistry》2009,107(1):112-122
The receptor tyrosine kinases ErbB2 and ErbB3 are phosphorylated in response to injury of the airway epithelium. Since we have shown that the membrane mucin MUC4 can act as a ligand/modulator for ErbB2, affecting its localization in polarized epithelial cells and its phosphorylation, we questioned whether Muc4 was involved, along with ErbB2 and ErbB3, in the damage response of airway epithelia. To test this hypothesis, we first examined the localization of MUC4 in human airway samples. Both immunocytochemistry and immunofluorescence showed a co‐localization of MUC4 and ErbB2 at the airway luminal surface. Sequential immunoprecipitation and immunoblotting from airway cells demonstrated that the MUC4 and ErbB2 are present as a complex in airway epithelial cells. To assess the participation of MUC4 in the damage response, cultures of NCI‐H292 or airway cells were scratch‐wounded, then analyzed for association of phospho‐ErbB2 and ‐ErbB3 with MUC4 by sequential immunoprecipitation and immunoblotting. Wounded cultures exhibited increased phosphorylation of both receptors in complex with MUC4. Scratch wounding also increased activation of the downstream pathway through Akt, as predicted from our previous studies on Muc4 effects on ErbB2 and ErbB3. The participation of MUC4 in the phosphorylation response was also indicated by siRNA repression of MUC4 expression, which resulted in diminution of the phosphorylation of ErbB2 and ErbB3. These studies provide a new model for the airway epithelial damage response, in which the MUC4–ErbB2 complex is a key element in the sensor mechanism and phosphorylation of the receptors. J. Cell. Biochem. 107: 112–122, 2009. © 2009 Wiley‐Liss, Inc. 相似文献
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Bovine substance P has been isolated in pure form from hypothalamic fragments and its complete amino acid sequence determined by studies performed on the intact peptide and on its isolated papain-generated fragments. Direct evidence for the positioning of each residue was obtained, amide assignments were unequivocally established, and the COOH-terminal residue was isolated and identified as Met-NH2. The results of total enzymic digestion performed on each of the peptides obtained argue against the presence of any non-amino acid constituents in the molecule. The amino acid sequence obtained is identical with that previously reported for material isolated form bovine colliculi and from equine small intestine. 相似文献
4.
Structure of a biologically active neurotensin-related peptide obtained from pepsin-treated albumin(s) 总被引:3,自引:0,他引:3
Using a radioimmunoassay toward the COOH-terminal region of neurotensin, an immunoreactive and biologically active neurotensin-related peptide (NRP) has been isolated from pepsin-treated fractions of bovine, canine, human, and rat plasma. Bovine NRP was identified as H-Ile-Ala-Arg-Arg-His-Pro-Tyr-Phe-Leu-OH, which is similar in structure to both neurotensin and angiotensin I. Canine and human NRP also had the above amino acid composition, whereas that obtained from rat plasma had valine substituted for isoleucine. At their concentrations in pepsin-treated plasmas (2-6 microM) rat, human and canine NRP were shown to increase vascular permeability when injected intradermally into rats and to release histamine from rat mast cells in vitro. The pure peptides also cross-reacted very effectively at nanomolar concentrations in a radioreceptor assay for neurotensin. The protein(s) which liberated NRP upon pepsin treatment were purified about 7-fold and shown to behave like albumin during sodium dodecyl sulfate-polyacrylamide gel electrophoresis, isoelectric focusing, and high pressure liquid chromatography on muBondapak C4. In addition, the purified preparations were found to react with anti-albumin antisera during immunodiffusion. Although the amino acid sequence of NRP was not found in albumin, a partial sequence homology was noted for NRP and various segments of bovine albumin. Using V8 protease, glutamyl residues were shown to lie within 3-4 amino acids of each end of NRP, as also occurs for the related segments in albumin. These results suggest that a subset of albumin-related protein(s) could serve as precursor(s) to biologically active neurotensin-related peptide(s). 相似文献
5.
Lysates of isolated rat polymorphonuclear leukocytes and macrophages were found to generate xenopsin-related peptides when incubated with a liver extract used as a source of precursor. The lysosomal enzyme, cathepsin D, was also shown to display this property and to share with the lysate a similar pH dependence (optimum, approximately pH 3.5) and sensitivity to the acid protease inhibitor, pepstatin A (ID50: lysate, 10 nM; cathepsin D, 30 nM). When subjected to HPLC on mu-Bondapak C-18, the xenopsin-related peptides generated by the lysate eluted near to those formed by cathepsin D and when tested in a radioreceptor assay for neurotensin, they displayed similar cross-reactivities (peak 2, approximately 50%; peak 1, approximately 100%). These results indicate that cathepsin D from lysed granulocytes can process precursor protein(s) to form radioreceptor-active xenopsin-related peptides. 相似文献
6.
R E Carraway 《The Journal of biological chemistry》1984,259(16):10328-10334
Using a radioimmunoassay directed toward the COOH-terminal, biologically active region of mammalian neurotensin (NT), the rapid (within seconds) generation of immunoreactive NT (iNT) during acid extraction of mammalian and avian gastric tissues has been demonstrated. Levels of iNT were shown to increase 25-200-fold in time. The reaction occurred in 0.1 N HCl and 2 N acetic acid and was prevented by raising the pH above 5 or by adding acetone. The temperature and pH dependence and the ability of pepstatin A to inhibit the reaction suggested the involvement of a pepsin-related acid protease. Furthermore, the reaction could be mimicked by incubating a stabilized gastric extract with hog pepsin at pH 2. Size exclusion chromatography demonstrated the presence of a precursor-like substance with an apparent Mr of 60,000. Although iNT generated in avian and mammalian gastric extracts could be distinguished chromatographically from NT in that species, the partially purified gastric iNT was active in a bioassay for NT which quantitates changes in vascular permeability after intradermal injection into rats. One might suggest that iNT serves as a signal within the gastric lumen, being generated at low pH by secreted pepsin. It is also possible that iNT could be formed within blood or gastric interstitial fluid by the action of pepsin-related (cathepsin or renin-like) enzymes at normal physiologic pH. 相似文献
7.
Levator advancement technique for eyelid ptosis 总被引:1,自引:0,他引:1
There have been many procedures advocated for the treatment of eyelid ptosis. The technique advocated in this paper consists of careful dissection and identification of anatomic landmarks, including preaponeurotic fat, Whitnall's superior transverse ligament, and the vertically oriented blood supply of the levator muscle. The attachment of the levator muscle into the cephalad portion of the levator muscle into the cephalad portion of the levator aponeurosis can be identified and easily dissected in order to perform the procedure of detachment and advancement to the tarsal plate. This procedure for ptosis has been successful in management in moderate to severe ptosis and in some cases has actually increased the muscle function, thereby enhancing the result. In this technique, the full length of levator muscle remains, so maximum excursion is achieved postoperatively. In addition, this surgical approach may be utilized for levator-lengthening procedures in cases of thyroid exophthalmus or overcorrected ptosis simply by performing the reverse procedure of detachment and insertion of a spacer based on the same ratio. Good results have been achieved in over 20 patients, with the exception of two patients who had absent to poor function and in whom undercorrection was present postoperatively. 相似文献
8.
Insertion mutations in the dam gene of Escherichia coli K-12 总被引:40,自引:0,他引:40
M G Marinus M Carraway A Z Frey L Brown J A Arraj 《Molecular & general genetics : MGG》1983,192(1-2):288-289
The dam gene of E. coli can be inactivated by insertion of Tn9 or Mud phage. Strains bearing these mutations are viable indicating that the dam gene product is dispensable. 相似文献
9.
Antisera towards neurotensin (NT) and the structurally related peptide, LANT6, were used to characterize immunoreactive peptides and proteins in extracts of chicken tissues. A 17 kDa protein was identified by Western blotting as a potential precursor to NT and LANT6. However, the posttranslational processing of this common precursor appeared to be tissue specific, giving rise to disproportionate amounts of NT and LANT6, along with varying expression of a large molecular LANT6 (Mr, 15 kDa). The intestinal cells containing immunoreactive NT, LANT6, and large molecular LANT6 behaved similarly during fractionation by size and density. These activities also banded together in particles resembling vesicles during centrifugation of isotonic homogenates of tissue. These results suggest that chicken NT and LANT6 are biosynthesized as parts of the same precursor, the processing of which can give rise to a variety of products stored within secretory vesicles. 相似文献
10.
C A Carothers Carraway H Fang X H Ye S H Juang Y C Liu M E Carvajal K L Carraway 《The Journal of biological chemistry》1991,266(24):16238-16246
[14C]Glucosamine metabolic labeling and concanavalin A blots were used to identify four major glycoprotein species associated with ascites tumor cell microvillar microfilament cores and with a transmembrane complex containing actin. Phalloidin shift analysis of glucosamine-labeled microvilli showed that glycoproteins of 110-120, 80, 65, and 55 kDa are stably associated with the microfilament cores. Analysis of large (greater than 10(6) kDa) transmembrane complexes from microvillar membranes made under microfilament-depolymerizing conditions (Carraway, C. A. C., Jung, G., and Carraway, K. L. (1983) Proc. Natl. Acad. Sci. U. S. A. 80, 430-434) revealed glycoproteins of the same Mr values, showing the same relative staining or labeling patterns as those observed with the microfilament cores. Gel filtration of high salt, high pH extracts of intact microvilli, microfilament cores, or transmembrane complexes showed that in all of these fractions the glycoproteins are associated in a very large, stable complex. The glycoprotein multimer was isolated essentially free of actin and other components by Sephacryl S-1000 chromatography of microvilli, microvillar membranes prepared at pH 11, microfilament cores, or transmembrane complex fractions in Triton X-100, 1 M KCl, glycine, pH 9.5. Purified glycoprotein complex bound actin when incubated under polymerizing conditions. The presence of the glycoprotein heteromultimer in both microfilament cores and transmembrane complex from isolated membranes and the association of the purified glycoprotein complex with actin are consistent with our hypothesis that the glycoprotein-containing transmembrane complex is an association site for microfilaments at the plasma membrane. 相似文献