Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: UMLS:C0272170 (SDS)
 50,377 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The production of enkephalin (EK) in the rat dental pulp was studied in pharmacological and biochemical aspects of EK-producing enzyme, EK precursor protein and the regulation of EK production. The EK precursor protein was primarily distributed in the microsomal fraction, and a common precursor protein (Mr about 58,000) was partially purified by Sephadex G-100 chromatography. Since the EK-producing enzyme, however, was mainly localized in the lysosomal fraction, and was found to be a cysteine proteinase, the lysosomal cysteine proteinases, cathepsins H, B and L, were separated by CM Sephadex C-50 ion exchange chromatography, and identified in respects to substrate specificity, pH optimum and inhibitor sensitivity. The EK-producing activity of the cathepsin B was demonstrated using the partially purified EK precursor protein from the pulp tissue as a substrate. The cathepsin B was further purified by Sephadex G-75 gel filtration to a 400-fold purity, and SDS polyacrylamide gel electrophoresis of the enzyme showed a distinct homogeneity (Mr about 23,600). The purified enzyme cleaved BAM-12P, a met-EK-containing peptide from bovine adrenal medulla, to met-EK-Arg6, but did not convert met-EK-Arg6 to met-EK, suggesting an endopeptidase activity of the enzyme. On the other hand, a concentration-dependent activation of the enzyme by bradykinin (BK) and des-Arg9-BK was found to be mediated through B1 receptor in intact pulp tissue. It was also demonstrated that intact structure of lysosomes and Ca++ were necessary for the activation of the enzyme by BK.
 ...
PMID:Pharmacological and biochemical study on the mechanism of enkephalin production in rat dental pulp. 213 Jan 76

The Semliki-Forest-virus-specific nonstructural proteins are translated as a large polyprotein (2431 amino acid residues), from which the mature polymerase components nsP1, nsP2, and nsP4 are released by proteolytic cleavages. The complete ns polyprotein (P1234) can be cleaved in two alternative ways yielding either P123 (with sequences of nsP1, nsP2 and nsP3) and nsP4 or P12 (nsP1 plus nsP2) and P34 (nsP3 plus nsP4). We studied the possible autoproteolytic role of nsP4 involved in the cleavage between nsP3 and nsP4 in an in vitro transcription-translation system. cDNAs encoding P34 precursor and shorter precursor protein segments covering the nsP3-nsP4 cleavage region, were cloned under the T7 RNA polymerase promoter. The mRNAs synthesized in vitro were capped and translated in rabbit reticulocyte lysates. The translational products were analyzed by SDS/PAGE. The precursor proteins containing nsP4 sequences were cleaved yielding the products with expected sizes, indicating that the cleavage took place at the nsP3-nsP4 junction. By deleting and truncating the cDNA coding for nsP4, the proteolytic activity was mapped within the 102 amino-terminal amino acids of nsP4. The cleavage between nsP3 and nsP4 can be inhibited by pepstatin A and probably takes place in cis, since exogenously added nsP4 was unable to mediate it.
 ...
PMID:The Semliki-Forest-virus-specific nonstructural protein nsP4 is an autoproteinase. 213 9

The cDNA sequence of the large dsRNA segment (segment A) of the N1 strain of infectious pancreatic necrosis virus (IPNV) has been determined. The nucleotide and deduced amino acid sequences were compared to the sequences of segment A of the Jasper strain of IPNV and to the sequences of segments A and B (5' and 3' flanking regions) of the 002-73 strain of infectious bursal disease virus (IBDV). The comparison demonstrated that the precursor protein of the major structural polypeptide, pVP2, is highly conserved at the N and C termini, whereas the amino acid sequence of an internal segment shows greater diversity between the strains. This internal segment probably carries the serotype-specific epitopes of birnaviruses. An alternative open reading frame (ORF) (444 bp) partly overlapping with the large ORF (2916 bp) of segment A was found to be conserved among the IPNV strains and is probably also present in the 002-73 strain of IBDV. This small ORF may encode a novel birnavirus polypeptide with an Mr of 17K. SDS-PAGE of radiolabelled purified IPNV particles revealed a band corresponding to the possible novel 17K polypeptide. Short terminal inverted repeats are found in segment A of the N1 and Jasper strains of IPNV and in segment B of the 002-73 strain of IBDV. Segment A of IPNV and segment B of IBDV also contain adjacent inverted repeats at their 5'-terminal flanking regions.
 ...
PMID:Sequence of the large double-stranded RNA segment of the N1 strain of infectious pancreatic necrosis virus: a comparison with other Birnaviridae. 230 63

Cellular membrane receptors for the immunostimulatory neuropeptide substance P have been previously identified on the cultured lymphoblast cell line, IM-9. The regulation of this receptor by ligand and the contribution to its molecular weight by N-linked sugars was studied by incubating IM-9 cells for 14 hr in the presence of [35S]met with or without substance P and tunicamycin, respectively. Cells were lysed and the receptor proteins were immunoprecipitated with an anti-receptor monoclonal antibody. SDS-PAGE analysis of untreated cellular lysates revealed specifically precipitated proteins of 38 kD and 33 kD, which were down-regulated by substance P. In tunicamycin-treated cells, whose substance P binding was not affected, the major immunoprecipitated protein had an apparent Mr of 29 kD. The time course of receptor processing was studied by pulse chase analysis. Three proteins of molecular weights 38 kD (mature receptor), 36 kD and 33 kD (receptor precursors) were identified for time periods of 30 min to 4 hr. The half life of the mature receptor and its precursors was approximately 1 hr and 0.5 hr, respectively. Results from the present studies suggest that the lymphocyte substance P receptor is translated as a precursor protein that is glycosylated.
 ...
PMID:Processing of the human IM-9 lymphoblast substance P receptor. Biosynthetic and degradation studies using a monoclonal anti-receptor antibody. 245 May 42

VP-2b, a major structural protein of infectious bursal disease virus (IBDV), and its precursor protein VP-2a were separated in a soluble form from the supernatant of ultracentrifuged viruses by using a monoclonal antibody specific for VP-3, the other major structural protein, to remove soluble VP-3 and remaining virus particles. The native VP-2a/2b inhibited the majority of virus-neutralizing (VN) activity in chicken anti-IBDV sera and chickens immunized with VP-2a/2b produced VN antibodies that passively protected susceptible chickens from infection. However, the separated VP2a/2b was not as immunogenic as intact virus particles. VP-2a/2b would appear to contain a conformational epitope which is destroyed by SDS and boiling and which may prove to be of critical importance in a subunit vaccine against type 1 IBDV.
 ...
PMID:A conformational immunogen on VP-2 of infectious bursal disease virus that induces virus-neutralizing antibodies that passively protect chickens. 254 87

Conformations of an artificial mitochondrial precursor protein pCox IV-DHFR have been analyzed by CD and fluorescence spectroscopy in the presence of (cardiolipin-rich) phospholipid vesicles or SDS micelles. Binding of pCox IV-DHFR to phospholipid vesicles involves a conformational change, which is presequence-dependent, accompanies alteration in the secondary structure of the DHFR moiety, but is different from total unfolding of the polypeptide chain. On the other hand, a conformational change of the fusion protein on binding to the micelles of a positively charged detergent, SDS, is not presequence-dependent.
 ...
PMID:Conformational changes of a mitochondrial precursor protein on binding to phospholipid vesicles and SDS micelles. A circular dichroism and fluorescence spectroscopy study. 266 Dec 63

The subcloning in pBR322 of the gene of the S. mutans OMZ 175 74K SR protein, was performed after in vitro reconstitution, from two recombinant EMBL3 phages, lambda SmAD9 and lambda SmAD10. The gene is expressed in E. coli HB101 under the control of its own promoter and produces a protein with a molecular weight of 195 kDa. A strong immunological relationship between the expressed protein and the 74K SR protein was observed in ELISA, Western blotting and immunoprecipitation. The 195 kDa protein was purified by immunoaffinity chromatography to homogeneity as judged by SDS-PAGE and native PAGE. Its reactivity with monoclonal anti 74K SR antibodies indicates that it is probably a precursor form of the 74K SR protein produced in S. mutans. The adhesion properties of the two proteins, tested in solid phase adherence assays, are quite similar. This indicates that the additional peptide present in the precursor protein has little or no role in the adherence properties of protein 74K SR.
 ...
PMID:Purification and characterization of the expression product of the sr gene of Streptococcus mutans OMZ 175. 266 61

A recombinant plasmid encompassing the human immunodeficiency virus type 1 (HIV 1) protease coding sequence and flanking regions (Ala-13 to Gly-185 of the pol open reading frame) has been expressed in two distinct strains of Escherichia coli, AR58 and AR68. In the first strain, AR58, the primary translation product, a 25 kilodalton (kDa) precursor protein, is short-lived and rapidly processes itself to the 11 kDa mature protease in vivo. In the second strain, AR68, the 25 kDa species is only partially processed, and it, a 13 kDa intermediate, and the mature 11 kDa enzyme accumulate at a ratio of 3:4.5:2.5, respectively. The 11 kDa mature protease from AR58 and the 25 kDa precursor from AR68 have been purified to homogeneity. The yield of 11 kDa enzyme from AR58 is approximately 0.02 mg/g wet weight of E. coli cell pellet. The protease has both the expected NH2- and COOH-terminal sequences. The yield of 25 kDa enzyme from AR68 is approximately 0.1 mg/g wet weight of E. coli cell pellet. In vitro, the 25 kDa precursor enzyme rapidly (t1/2 approximately equal to 9 min) processes itself into a species with a mass of approximately 13 kDa and a species with a mass of approximately 11 kDa. Both of these latter species can be separated by RP-HPLC, have the NH2-terminal sequence expected for the mature protease, and are active. The 11 kDa enzyme from AR58 comigrates with the 11 kDa enzyme from AR68 on RP-HPLC and SDS polyacrylamide gel electrophoresis. On extended incubation at 4 degrees C at either neutral or acidic pH all species of the protein exhibit further autodegradation at defined sequences. The availability of the mature, 11 kDa enzyme and the 25 kDa precursor will allow biochemical and physical studies on this critical viral enzyme.
 ...
PMID:Characterization and autoprocessing of precursor and mature forms of human immunodeficiency virus type 1 (HIV 1) protease purified from Escherichia coli. 269 27

Saccharomyces cerevisiae harboring linear dsDNA plasmids, pGKL1 and pGKL2, secretes a killer toxin consisting of 97, 31 and 28 kilodalton subunits (Nucleic Acids Res., 15, 1031-1046, 1987). We isolated the DNA encoding the N-terminal pre-sequence of the 28K precursor protein and constructed a new secretion vector in S. cerevisiae. Mouse alpha-amylase fused to the 28K signal sequence was secreted into the culture medium with a high efficiency similar to those fused to the mating factor alpha and 97K-31K killer signal sequences. This data clearly indicates that 28K presequence functions as a secretion signal. Glycosylated and nonglycosylated alpha-amylase molecules were detected in the culture medium. The secretion of alpha-amylase was blocked by sec18-1 mutation. The secreted alpha-amylase recovered from the medium was found to migrate faster in SDS-polyacrylamide gel than the precursor form of alpha-amylase synthesized in vitro. These lines of evidence suggest that mouse alpha-amylase fused to 28K killer signal sequence was processed, glycosylated and secreted through the normal secretion pathway of the yeast.
 ...
PMID:A novel yeast secretion signal isolated from 28K killer precursor protein encoded on the linear DNA plasmid pGKL1. 304 59

A polyclonal antiserum was prepared in rabbits against the structural glycoprotein (SGP) complex previously isolated from a bacterial collagenase digest of bovine corneal stroma (R. Alper, Curr. Eye Res. 2:479, 1983). Direct and indirect enzyme-linked immunosorbent assays indicated that the antiserum was specific for the SGP-complex and did not react with Types I, III and IV collagen, fibronectin, laminin or actin. Immunoblot experiments indicated that the antiserum reacted with all of the components of the SGP-complex as well as with the cell matrix laid down by bovine keratocytes in culture. An attempt was made to isolate individual antibodies from the antiserum by selective elution from immunoblots of the components of the SGP-complex separated by SDS-PAGE. It was found that regardless of the protein band from which the antibody was eluted, every antibody isolated reacted with every protein component of the SGP-complex suggesting that the SGP-complex may have been derived from a single precursor protein and that the observed heterogeneity of the SGP-complex may have been the result of proteolytic breakdown of the protein held together by disulfide bonds. When the anti-SGP antiserum was used to immunoprecipitate 14C-proline labeled proteins from the media of bovine keratocytes in culture, the major protein observed had a Mr of about 140,000 daltons, similar to that of GP-140 also known as CL-glycoprotein. These proteins have been shown to represent the tissue form of Type VI collagen. To test the hypothesis that the SGP-complex may be related to the GP-140 (CL-glycoprotein), ELISA and immunoblotting studies were performed comparing the properties of the anti-SGP serum with those of a polyclonal antibody specific for Type VI collagen. The SGP-complex reacted positively by ELISA with the anti-human Type VI collagen antiserum and, conversely, human Type VI collagen gave a positive ELISA reaction with an antiserum against the SGP-complex. The anti-human Type VI collagen antiserum reacted with most of the major components of the SGP-complex on immunoblots of SDS-PAGE gels. These data indicate that the SGP-complex is related to and probably is derived from the tissue form of Type VI collagen.
 ...
PMID:The bovine corneal SGP-complex is related to the tissue form of type VI collagen. 335 3


<< Previous 1 2 3 4 5 6 7 8 9 10 Next >>