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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A convenient method for the determination of unfolding rates of small globular proteins under physiological conditions was developed using digestion with proteases. The apparent first-order rate constants for digestion of lysozyme with thermolysin and with Pronase at pH 8 and 50 degrees C were shown to be saturated with increases of concentrations of these proteases. The maximum rate constants extrapolated were identical in digestions with two different proteases, and were found to be equal to the unfolding rate constant of lysozyme. Similarly, the unfolding rate constant of RNase A at pH 8 and 50 degrees C, and those of lysozyme, RNase A and beta-lactoglobulin at pH 8 and 40 degrees C, were determined by the digestion method. Thus, it was shown that digestion by proteases proceeds mainly via the unfolded state of proteins.
J Mol Biol 1986 Aug 20
PMID:Unfolding rates of globular proteins determined by kinetics of proteolysis. 378 15

A group of closely related high mol. wt (Mr) membrane glycoproteins is expressed with varying Mr on different subpopulations of lymphocytes, but the different Mr forms share the Ly-5 and T200 antigenic determinants. The Ly-5 molecule expressed by thymocytes has an Mr of 175,000 (175ly5). The antigenically related molecule on B-cells has an Mr of 210,000 (210ly5). It is not known whether the variations in size are due to differences in the polypeptide chain, post-translational modifications such as glycosylation, or both. In this report we examine the glycosylation of 175ly5 and 210ly5 to determine whether differences in carbohydrate moieties may account for the different Mr of these two Ly-5 species. Pronase digestion and alkaline borohydride treatment of these molecules labeled in the terminal galactose residues revealed that 210ly5 molecules have a more complex oligosaccharide pattern than 175ly5 molecules. While Ly-5 oligosaccharides from a T-cell tumor line were very similar to those of normal thymocytes, the pattern of Ly-5 carbohydrates from a B-cell tumor were somewhat different than those from normal B-cells. This report also presents evidence for O-linked sugars on Ly-5 molecules.
Mol Immunol 1985 May
PMID:Analysis of structural differences between Ly-5 molecules of T- and B-cells. 387 17

Gunn rats have a marked deficiency in hepatic UDP-glucuronosyl transferase activity which results in hyperthyroxinemia and hyperbilirubinemia. Their thyroids show a brownish-black discoloration associated ultrastructurally with intracellular dense granules and intraluminal dense masses. In order to determine whether colloid composition and colloid proteolysis are altered in the thyroid of the Gunn rat compared with the Wistar rat, we studied the in situ resistance of thyroid proteins to in vitro proteolysis, the pattern of in vivo (125I) labeled thyroid iodoproteins and the proteolysis of isolated iodoprotein fractions in both strains of rats. For the cytochemical study, thin sections of aldehyde-fixed and plastic-embedded thyroid tissue were treated with 0.3 or 1% pronase in aqueous solution. With the low concentration of pronase, the secretory granules in C-cells and the apical vesicles in follicular cells were extensively digested in both strains of rats, whereas the colloid in the follicular lumen and the colloid droplets were only partially digested. With the high concentration of pronase, the colloid in the lumen and the colloid droplets were more markedly digested in both strains. In the presence of both concentrations of pronase, the dense granules and intraluminal dense masses were unchanged in the Gunn rats. The (125I) iodoprotein pattern was investigated 24 h after a single injection of (125I) iodide and by labeling at the isotopic equilibrium. It was found that the (125I) thyroglobulin fraction was reduced, whereas the (125I) 3-8 S fraction was increased in Gunn rats compared to Wistar rats. Pronase hydrolysis of the soluble (125I) iodine fraction showed similar pronase-resistant fractions in both strains with the single labeling procedure. At the isotopic equilibrium, the pronase resistant fraction was significantly increased in Gunn rats (Gunn 24.0 +/- 5.3%; Wistar: 13.7 +/- 3.1% of the soluble 125I) and a linear correlation was observed between the (125I) 3-8 S fraction of the soluble extract and the pronase-resistant fraction. These data suggest that iodocompounds of small molecular size and low turnover accumulate in the thyroid of the Gunn rat due to their strong resistance to in vivo hydrolysis. A local accumulation of 3-8 S iodocompounds may occur within the intracellular dense granules and intraluminal dense masses in the thyroid of Gunn rat.
Virchows Arch B Cell Pathol Incl Mol Pathol 1983
PMID:Abnormal iodoprotein distribution and resistance to proteolysis in Gunn rat black thyroid. An ultrastructural and biochemical study. 614 65

Normal fecal antigen-1 (NFA-1), which is a carcinoembryonic antigen (CEA)-related glycoprotein with a mol. wt of 20,000-30,000, was purified from normal adult feces by immuno-adsorption, gel filtration and ion exchange chromatography. Highly purified NFA-1 was partially cross-reactive with CEA but antigenically unrelated to nonspecific cross-reacting antigen (NCA) which was also cross-reactive with CEA. NFA-1 also had a unique determinant not present in CEA or other related antigens including NCA. In a solid-phase RIA system, the reactivity of NFA-1 with a specific anti-CEA antiserum was much stronger than that of NCA. Although digestion with Pronase E did not affect the antigenicity of NFA-1, reduction and alkylation destroyed its antigenic reactivity. The total amount of carbohydrate in NFA-1 was 13.3%, compared to 52.4% in CEA and 21.6% in NCA. The amino acid composition of NFA-1 was similar to that of CEA. The sequence of the first 10 NH2-terminal amino acids in NFA-1 was Ala-Glu-Pro-Pro-Lys-Pro-Phe-Ile-(Thr)-Ser. This was totally different from that of the first NH2-terminal amino acids of CEA isolated from tumor tissue.
Mol Immunol 1982 Mar
PMID:Immunological characterization and structural studies of normal fecal antigen-1 related to carcinoembryonic antigen. 617 63

Polypeptides co-purifying with DNA in alkali are covalently bound to DNA. DNA purified by treatment with alkali, sodium dodecyl sulphate and phenol absorbed 125I under conditions designed to radioiodinate exclusively tyrosine and histidine in peptides. A significant amount of the absorbed 125I remained associated with DNA during treatment with phenol as well as during precipitation with ethanol from neutral and alkaline solutions. However, after prolonged digestion with proteinase K, most of the radiolabelled material could be removed from 125I-treated DNA. Further treatment with a second protease (Pronase) released no larger fraction of the 125I label. The residual radiolabelled material could be precipitated together with DNA by ethanol and it remained associated with DNA also in the presence of alkali (95 degrees C), acid (37 degrees C) and hydroxylamine (37 degrees C). In contrast, radiolabelled peptides were released from DNA by treatment with hot piperidine (10% at 95 degrees C) and by agents that hydrolyse peptides and modify DNA, e.g. strong acid (95 degrees C) and formic acid/diphenylamine. The radiolabelled peptides, once released from DNA by these chemical methods, could be further cleaved by Pronase. This shows that the residual DNA/peptide complex isolated after prolonged protease digestion is protease-resistant unless it is cleaved or otherwise modified by harsh chemical treatment. The linking groups between deoxynucleotides and the radiolabelled residual peptides could be isolated by digestion of DNA in the DNA/peptide complex. Radiolabelled peptides could be released from this linking group material by phosphodiesterases, indicating the involvement of phosphodiesters in the linking groups.
J Mol Biol 1983 Feb 25
PMID:Phosphodiester bonds between polypeptides and chromosomal DNA. 630 72

Upon superinfection of immune (lysogenic) cells with bacteriophage Mu, a form of Mu DNA accumulates that sediments about twice as fast as the linear phage DNA marker in neutral sucrose gradients. This form is also detected upon infection of sensitive cells with Mu. We have purified it and examined its physical nature. Under the electron microscope it appears circular and supertwisted. Upon treatment with Pronase, phenol or sodium dodecyl sulfate, however, it is converted to a linear Mu-length form, indicating that the circle is not covalently closed. The linear DNA still has heterogeneous host sequences at its termini. The circular DNA is resistant to the action of Escherichia coli exonuclease III and T7 exonuclease, but becomes sensitive to these nucleases after treatment with Pronase showing the presence of a protein that binds non-covalently to the ends of the DNA to circularize it as well as protect it from digestion with exonucleases. The complex is resistant to high salt (up to 6 M-NaCl) but can undergo transitions between forms that are partially open, open circular, linear and circular dimers and trimers. Examination of DNA from mature phage particles reveals that a circular DNA species is present in at least 0.1 to 1% of the population. The purified complex is extremely efficient in transfection of E. coli spheroplasts. We estimate the molecular weight of the protein in this DNA-protein complex to be approximately 64,000, and suggest that this complex might represent the integrative precursor of infecting Mu DNA.
J Mol Biol 1983 Jun 25
PMID:Infecting bacteriophage mu DNA forms a circular DNA-protein complex. 630 60

The major polypeptides composing hepatitis B surface antigen (HBsAg) particles are P-I and P-II. P-II shares the same amino acid sequence as P-I and contains an additional carbohydrate moiety of mol. wt approximately 5000. When a purified preparation of P-II was digested with Nagarse and then with Pronase P, it gave rise to a glycopeptide containing 15 amino acid residues and the carbohydrate moiety of P-II. The N-terminal amino acid sequence of the glycopeptide was determined to be Lys-Pro-Thr-Asp-Gly-Asn-. The polysaccharide moiety contained 5 moles of N-acetylglucosamine and was connected with Asn at the sixth position from the N-terminus. When mice were immunized against this HBsAg glycopeptide, they raised humoral antibodies which bound to each of three preparations of P-I derived from HGsAg particles of subtypes adw, adr and ayw, thereby indicating that the sequence of 15 amino acids in the glycopeptide would constitute a common antigenic structure of HBsAg.
Mol Immunol 1982 Sep
PMID:A glycopeptide containing 15 amino acid residues derived from hepatitis B surface antigen particles: demonstration of immunogenicity to raise anti-HBs in mice. 714 54

Streptomyces griseus excretes a small molecular mass (30 kDa) aminopeptidase that could be used for various biotechnological applications. This enzyme was isolated from an extracellular protease mixture of Streptomyces griseus (Pronase E. Sigma) and single crystals were obtained by the vapor diffusion method using polyethylene glycol 4000 as the precipitant. The crystals belong to the tetragonal space group P4(1)2(1)2 (P4(3)2(1)2), with cell dimensions of a = b = 61.82(3) A and c = 145.88(4) A. These crystals are mechanically strong, they are stable in the X-ray beam and they diffract to better than 1.8 A resolution. The cell dimensions and the cell symmetry are consistent with one molecule in the asymmetric unit and the crystals are suitable for a detailed high-resolution crystallographic analysis. A complete native data set to 1.9 A resolution has been collected on a Rigaku R-AXIS-IIC Imaging Plate Detector system and a heavy-atom derivative search is in progress.
J Mol Biol 1993 Mar 05
PMID:Crystallization and preliminary crystallographic analysis of Streptomyces griseus aminopeptidase. 845 May 45

Extracts of human liver were found to contain activities which copurified and coeluted with the two major subtypes of mediators (type A and type P) isolated from insulin-stimulated rat liver. The putative type A mediator from human liver inhibited cAMP-dependent protein kinase from bovine heart, decreased phosphoenolypyruvate carboxykinase mRNA levels in rat hepatoma cells, and stimulated lipogenesis in rat adipocytes. The putative type P mediator stimulated bovine heart pyruvate dehydrogenase phosphatase. Both fractions were able to stimulate proliferation of EGFR T17 fibroblasts and the type A was able to support growth in organotypic cultures of chicken embryo cochleovestibular ganglia. Both activities were resistant to Pronase treatment and the presence of carbohydrates, phosphate, and free-amino groups were confirmed in the two fractions. These properties are consistent with the structure/ function characteristics of the type A and P inositolphosphoglycans (IPG) previously characterized from rat liver. Further, the ability of the human-derived mediators to interact with rat adipocytes and bovine-derived metabolic enzymes suggests similarity in structure between the mediators purified from different species. Galactose oxidase-susceptible membrane-associated glycosylphosphatidylinositols (GPI) have been proposed to be the precursors of IPG. GPI was purified from human liver membranes followed by treatment with galactose oxidase and reduction with NaB3H4. Serial t.l.c. revealed three radiolabeled bands which comigrated with the putative GPI precursors found in rat liver. These galactose-oxidase-reactive lipidic compounds, however, were only partially susceptible to hydrolysis with phosphatidylinositol-specific phospholipase C from Bacillus thuringiensis and were resistant to glycosylphosphatidylinositol-specific phospholipase C from Trypanosoma brucei. These data indicate that IPG molecules with insulin-like biological activities are present in human liver.
Biochem Mol Med 1997 Aug
PMID:Isolation and partial characterisation of insulin-mimetic inositol phosphoglycans from human liver. 925 87

The source and nature of zona lytic factors during zona escape of hamster blastocyts were investigated. When cultured in hamster embryo culture medium (HECM)-2h, all 8-cell embryos (n = 135) developed to zona escaped-blastocysts with complete zona lysis. In addition, 2-cell embryos, when co-cultured with zona escaping-blastocysts (at a ratio of 1:10), exhibited zona lysis. Various other embryos at the 1-8-cell stages also showed zona lysis when cultured with zona-escaping blastocysts. However, zonae from mice, rats, sheep and humans were resistant to lysis under these conditions. Pronase treatment resulted in rapid zona lysis in hamsters (7 +/- 1 s), whereas in other species zona lysis was much slower: mouse (662 +/- 27 s), rat (532 +/- 16 s), sheep (120 +/- 12 s) and human (104 +/- 8 s). When cysteine protease inhibitors (antipain, leupeptin, E-64 and p-hydromercuricbenzoate) were tested, they completely inhibited zona escape, while trypsin inhibitors (TLCK and SBTI) did not. Uterine zona lysin contribution in zona escape was discounted since: (i) uterine luminal flushing and endometrial extract from day 4 (the time of zona escape in vivo) pregnant females failed to lyse zonae and (ii) endogenous oocytes and transferred 2-cell embryos (to day 3 pseudopregnant recipients) were all zona-intact, while 71% of transferred blastocysts exhibited zona escape, following their recovery after 24 h. These observations suggest that a species-specific, embryonic proteolytic factor, with a cysteine protease-like activity, is involved in the zona escape of blastocysts in hamsters.
Mol Hum Reprod 2000 Nov
PMID:Evidence for the involvement of a species-specific embryonic protease in zona escape of hamster blastocysts. 1104 63


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