Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.27.4 (ribonuclease)
 6,621 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Dipolid human fibroblast-rich tissues contain a macromolecule with a molecular weight between 30,000 and 50,000 daltons which will inhibit the proliferation of fibroblasts in the G1 phase of the cell cycle (i.e., inhibit both 3H-thymidine uptake as well as the normal increase in cell number). The inhibitor is destroyed by trypsin but not by ribonuclease or deoxyribonuclease, and it is thermolabile. It has an acid IEP. It is not cytotoxic, and its inhibitory activity appears to be completely reversible. This fibroblast endogenous inhibitor does not interfere with the proliferation of DNA synthesis by human lymphocytes, bronchial carcinoma cells, or HeLa cells. The activity does not appear to be species specific. Therefore, we suggest that it is quite possible that the control of fibroblast proliferation resides in a fibroblast chalone. Diploid human fibroblasts, in contrast to chicken or mouse fibroblasts or heteroploid fibroblasts in general, stringently require serum for their proliferation. All of this mitogenic activity of calf serum can be concentrated in a molecular weight range around 100,000 daltons by ultrafiltration. All of the mitogenic activity within this molecular weight class can be concentrated at a pH of 5.2 via isoelectric focusing, and all of the activity at this isoelectric point can be concentrated in one peak on preparative polyacrylamide gel electrophoresis. This latter material is homogeneous at three different pH's in analytical gel electrophoresis as well as in SDS electrophoresis. This purified serum mitogen for diploid human fibroblasts in vitro also works in vivo and represents as much as 0.5% of calf serum protein, albeit there is much less of this protein in adult cow or horse. It is composed of two equal subunits weighing about 60,000 daltons each and contains about 2 moles of sialic acid, one S-S bond, and 6 moles of hexose per subunit. There is a reciprocal relationship between the biological activity of fibroblast inhibitor and serum mitogen, but there is no apparent direct interaction between these two proteins. Addition of pure serum mitogen to diploid human fibroblasts in vitro results in the release of commensurable chalone activity into the medium and a reciprocal loss of mitogen from the medium. Therefore, we propose that serum contains a single macromolecule which competes with endogenous chalone on the surface of diploid human fibroblasts and that this functions as an anti-chalone for the fibroblast.
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PMID:Circulating factors controlling cell proliferation. 13 64

The addition of trypsin [EC 3.4.21.4]-digested liver microsimes induced cyanideinsensitive respiration in guinea pig polymorphonuclear leucocytes with concomitant acceleration of the hexose monophosphate oxidative pathway. The respiration was insensitive to inhibitors of mitochondrial respiration but sensitive to glycolytic inhibitors. These metabolic alterations are similar to those associated with phagocytosis, though the digested mocrosomes were apparently not taken up by the cells and prpbably trigger the netabolic changes by interaction with the cellular membrane. Intact microsomes or microsomes treated with chymotrypsin [EC 3.4.21.1], bacterial proteinase, ribonuclease [EC 3.1.4.22], or neuraminidase [EC 3.2.1.18] could not induce such respiration.
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PMID:Metabolic pattern of polymorphonuclear leucocytes induced by trypsin-digested microsomes. 115 Jun 33

A fast atom bombardment mass spectrometric protocol has been developed to determine the type of oligosaccharide chain present in glycoproteins. The procedure is based on acetolysis of the intact glycoconjugate, extraction of the peracetylated carbohydrate fragments and analysis by fast atom bombardment mass spectrometry. The molecular ions present in the FAB spectra uniquely define the composition of the oligosaccharides with respect to hexose, aminohexose and sialic acid content. High mannose oligosaccharides yield a series of peracetylated hexose oligomers whereas complex-type oligosaccharides afford a series of N-acetyl-lactosamine containing species. Fucosylation is usually not detected but sialylated oligosaccharides are readily identified and the type of sialic acid is also defined. The method has been tested on three glycoproteins of known structure - fetuin, ribonuclease B and erythrocyte Band 3 - and on a glycoprotein of unknown structure - alpha-galactosidase I, an enzyme lectin from Vicia faba. The latter is shown to contain high mannose carbohydrate chains.
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PMID:A novel mass spectrometric procedure for the rapid determination of the types of carbohydrate chains present in glycoproteins: application to alpha-galactosidase I from Vicia faba seeds. 241 21

A new method is described for locating the specific sites of attachment of Asn-linked carbohydrates in glycoproteins. The molecular weights of peptides released from the glycoprotein with proteases of known specificity are determined by fast atom bombardment mass spectrometry and fitted to the known or DNA-derived sequence. Oligosaccharides attached to Asn are released either before or after proteolysis with a glycosidase, usually peptide: N-glycosidase F, an enzyme that cleaves the beta-aspartylglycosylamine linkage of all known types of Asn-linked sugars and converts the attachment-site Asn to Asp. New peaks appearing in the mass spectra after treatment with glycosidase correspond to formerly glycosylated sites. Conversely, signals which disappear after glycosidase treatment correspond to glycopeptides. The differences in mass between these sets of signals define the composition of the carbohydrate at the given site in terms of deoxyhexose, hexose, N-acetylhexosamine, and sialic acid content. The extent of glycosylation at a given site can be estimated from the ratio of the peak heights corresponding to the Asn- vs Asp-containing peptides which differ by 1 Da in mass. This rapid and sensitive (low nmol) technique is illustrated here for ribonuclease B and for tissue plasminogen activator, a multiply glycosylated glycoprotein.
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PMID:Carbohydrate mapping by mass spectrometry: a novel method for identifying attachment sites of Asn-linked sugars in glycoproteins. 309 66

Pyrophosphate, p-nitrophenyl phosphate and a variety of pyrimidine and purine nucleotides are hydrolyzed by the solubilized membrane-bound enzymes of the brush border plasma membrane of Hymenolepis diminuta. The pH optima (or ranges) for hydrolysis of substrates are 8.0 (pyrophosphate), 8.8 (p-nitrophenyl phosphate), 8.4-8.9 (nucleoside monophosphates), and 7.1-8.1 (nucleoside triphosphates); all substrates, with the exception of nucleoside triphosphates, have a higher affinity for the solubilized enzyme at pH 7.4 than at their optimal pH for hydrolysis. ATP is degraded completely by the enzyme preparation to adenosine and inorganic phosphate, but since neither ADP nor ATP accumulate in the incubation medium it is not known whether ATP hydrolysis involves the sequential hydrolysis of terminal phosphate groups. Isoelectric focusing and various chromatographic procedures (gel permeation, ion-exchange and hydrophobic interaction chromatography) fail to separate the alkaline phosphatase, phosphodiesterase, 5'-nucleotidase, adenosine triphosphatase and ribonuclease activities associated with the solubilized membrane preparation. Additionally, inhibitor studies indicate that only a single enzyme with low substrate specificity is involved in the hydrolysis of nucleotides, p-nitrophenyl phosphate, pyrophosphate and hexose phosphate esters. Purines and pyrimidines and their nucleosides interact with the active site, and in some instances activity of the enzyme is stimulated by an unknown mechanism.
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PMID:Nucleotide hydrolysis by solubilized membrane-bound enzymes of the brush border plasma membrane of Hymenolepis diminuta. 613 88

Glycosylation is one of the most important posttranslational modifications affecting the functions of proteins and cell activities. Mass spectrometry (MS) has proven to be an effective tool for structural glycobiology and has helped gain an understanding of glycoprotein-mediated diseases. Although electro-spray ionization-tandem MS remains widely recognized as an effective means for oligosaccharide characterization, the hydrophilic nature of glycans has often caused the poor ionization efficiency requiring either derivatization or nanoelectrospray to improve detection sensitivity. In this report we describe the use of a chip-based infusion nanoelectrospray platform coupled with the hybrid triple quadrupole/linear ion trap for identification and characterization of glycosylation in complex mixtures. The high-mannose-type N-glycosylation in ribonuclease B was used to map the glycosylation site and obtain glycan structures. Using the chip-based nanoelectro-spray with precursor ion scanning linear ion trap MS, we were able to map the glycosylation site and obtain the glycan structures in ribonuclease B at 100 fmol/microL in a single analysis. In addition, a new, low-abundant glycoform with an additional hexose (Hex10GlcNAc2) attached to ribonuclease B was discovered. The results reported here demonstrate that the chip-based infusion nanoelectrospray ionization coupled to a quadrupole/linear ion trap platform is a valuable system, as it provides high sensitivity and stability for nanoelectrospray analysis, and allows extended acquisition time for completing precursor ion scanning and subsequent MS2 and MS3 information in a single analysis.
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PMID:Characterization of protein glycosylation using chip-based nanoelectrospray with precursor ion scanning quadrupole linear ion trap mass spectrometry. 1646 44

A subcellular extract prepared from nonencapsulated Diplococcus pneumoniae type 3 protected mice against a subsequent challenge with virulent D. pneumoniae types 1, 2, 3, and 7. Potency ratios ranged from 25 (type 3) to >1,175 (type 1). The immunity induced in mice by this preparation was best obtained by intraperitoneal inoculation followed by intravenous challenge. Mice immunized in this manner were protected up to 12 weeks and could withstand a challenge of several hundred LD(50). The protection was destroyed by treatment of the preparation with ribonuclease and was decreased by treatment with protease. The preparation consisted of 60.5% ribonucleic acid, 29.1% protein, 6.3% deoxyribonucleic acid, and 4.0% hexose. Ultracentrifugation studies indicated that this extract had five constituents which are compatible with ribosomal material.
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PMID:Protection against pneumococcal infection by a ribosomal preparation. 1655 34

Tandem mass spectrometry of glycans and glycoconjugates in protonated form is known to result in rearrangement reactions leading to internal residue loss. Here we studied the occurrence of hexose rearrangements in tandem mass spectrometry of N-glycopeptides and reductively aminated N-glycans by MALDI-TOF/TOF-MS/MS and ESI-ion trap-MS/MS. Fragmentation of proton adducts of oligomannosidic N-glycans of ribonuclease B that were labeled with 2-aminobenzamide and 2-aminobenzoic acid resulted in transfer of one to five hexose residues to the fluorescently tagged innermost N-acetylglucosamine. Glycopeptides from various biological sources with oligomannosidic glycans were likewise shown to undergo hexose rearrangement reactions, resulting in chitobiose cleavage products that have acquired one or two hexose moieties. Tryptic immunoglobulin G Fc-glycopeptides with biantennary N-glycans likewise showed hexose rearrangements resulting in hexose transfer to the peptide moiety retaining the innermost N-acetylglucosamine. Thus, as a general phenomenon, tandem mass spectrometry of reductively aminated glycans as well as glycopeptides may result in hexose rearrangements. This characteristic of glycopeptide MS/MS has to be considered when developing tools for de novo glycopeptide structural analysis.
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PMID:Hexose rearrangements upon fragmentation of N-glycopeptides and reductively aminated N-glycans. 1941 47