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

Lactoferrin (LF) binding to the surface of human neutrophils was shown to be dependent upon the presence of cell surface DNA by (i) an abrogation of LF binding after treatment of whole cells with DNAse; (ii) an abrogation of LF binding to a purified cell membrane suspension after DNAse digestion, (iii) a restoration of LF binding, after initial treatment of cells with DNAse, by the addition of exogenous DNA. Using a biotinylated LF probe, no other binding molecules were found after SDS PAGE of neutrophil cell membrane proteins. Further evidence of a DNA-LF interaction was obtained by the co-isolation of LF with DNA by both gel chromatography and affinity chromatography using Heparin Sepharose CL 6B. The interaction of LF with neutrophils was a saturable phenomenon with a Kd of 6.2 X 10(-6) M and a maximum binding of 9.2 X 10(6) molecules per cell. These results suggest that cell membrane DNA may have a novel role as a receptor for LF, and indicates the need for further experiments to determine whether the functional effects of LF are modified by the DNAse treatment of LF responsive cells.
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PMID:Lactoferrin binds to neutrophilic membrane DNA. 370 57

A preparative, sequential chromatographic procedure has been developed for the purification of human gamma interferon (HuIFN-gamma). The four steps in the procedure are Controlled Pore Glass-adsorption chromatography, Concanavalin-A affinity chromatography, Heparin-Sepharose affinity chromatography and gel-filtration. By virtue of the development of a coordinated effluent-affluent buffer scheme, eluants can also serve as loading buffers for the succeeding column. Consequently, crude HuIFN-gamma preparations can be purified rapidly (approximately one week), easily, and is amenable to a semi-automated process. The procedure has also been shown to be efficient. Here, as an example, it is reported that an overall purification of greater than 75,000-fold can be achieved, yielding a specific activity of 5.2 X 10(7) units/mg, and a recovery of 95.5%. In addition, the peak fraction, representing 37.8% of the applied activity, had a specific activity of 1.0 X 10(8) units/mg protein and represents a purification of more than 145,000-fold. An SDS-PAGE analysis of one such fraction indicated that approximately 40% of the final material was HuIFN-gamma.
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PMID:A simple and efficient method for the purification of human gamma interferon. 641 49

Heparin treated and aldehyde crosslinked rat aorta segments were implanted in infrarenal aorta of homologous rats. One year following aortic replacement, the subendothelial scar and the prosthetic remnants were excised. The scar and the host intima-media were incubated with 3H-valine for 4 h and extracted with 5 M guanidinium chloride--0.05 M dithiothreitol--0.1 M Tris--0.1% EDTANa2 at pH 7.5 prior (Extract 1) and following (Extract 2) hydrolysis of collagen. The radioactivity of extract 1 accounted for approximately 80% of the total label incorporated in the scar and host intima-media. The 3H-label of extract 1 adjusted for the tissue collagen content was about twenty times higher in the scar than in the host aorta. The major 3H protein peaks from Extract 1 of scar and host aorta were of 130 K, 100 K and 70 K apparent molecular weight, based on polyacrylamide gel electrophoresis in SDS. Hydrolysis with 2N KOH of the extraction residue from the host aorta and scar yielded 3H-val-pro dipeptides and hydrolysis with 6N HCl desmosines. The incorporation pattern of 3H-valine into proteins and the presence of elastin synthesized de novo in the scar replacing the prosthesis indicate macromolecular repair of the host aortic wall.
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PMID:Healing of biodegradable vascular prosthesis. Incorporation of 3H-valine into proteins in the subendothelial scar and host intima-media of rat aorta. 667 80

Heparin binding to serum proteins and their subsequent precipitation is reportedly increased in cystic fibrosis (CF). We have confirmed this finding for CF patients over the age of 12 [11.34 +/- 2.42 mg/ml precipitated protein for normals (n = 19) versus 17.46 +/- 4.60 mg/ml for CF patients (n = 37), P less than 0.001; 0.629 +/- 0.098 mg/ml precipitated heparin for normals versus 0.789 +/- 0.206 for CF patients, P less than 0.01]. We have also shown that patients with a variety of pulmonary diseases unrelated to CF do not show this effect. When the amounts of protein and heparin precipitated are compared with the amount of IgG found in the whole serum sample, the correlation coefficients (protein r = 0.77; heparin, 0.74; n = 81) are significant at a level of P less than 0.001. In addition, the report that young CF patients exhibit hypogammaglobulinemia prompted us to examine serum samples from CF patients and age-matched controls under the age of 12. No differences were found. To investigate the molecular basis for this effect, sera from patients with CF and from age-matched controls were precipitated with 50 mg% heparin at pH 5.57. Pellets resolubilized in 8 M urea were fractionated on DEAE-Sephadex and analyzed by double-immunodiffusion, SDS-PAGE, immunoelectrophoresis, and radial immunodiffusion. IgG constituted 55-56% of the eluted protein. When serum from all donors was fractionated by Staph A-Sepharose into IgG and non-IgG fractions, 85-89% of heparin precipitable protein was in the IgG fraction.
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PMID:Identification of a major heparin-precipitable protein in human serum and its relationship to cystic fibrosis. 706 73

A 29-yr-old white female has suffered from recurrent venous thromboses over the last 12 yr. Plasma antithrombin III (AT-III) levels were 48% of normal by immunoelectrophoresis and 56% by chromogenic assay. Three of four siblings and the father had similar AT-III levels without associated venous thromboses. Heparin-Sepharose chromatography demonstrated normal behavior of the patient's AT-III. Her purified AT-III could not be distinguished from AT-III purified from a normal control either by SDS polyacrylamide gel electrophoresis or by crossed immunoelectrophoresis, and the heparin cofactor activity and the progressive antithrombin activity of both AT-III samples were identical. Turnover studies were made in the patient using her own purified AT-III labeled with 131I, (*I). The results did not differ significantly from studies made with autologous *I-AT-III in two normal control women. Her fractional breakdown rate of 0.54 total plasma AT-III per day compared with 0.45 and 0.52 in the controls. These studies indicate that the patient synthesizes a normal AT-III molecule at half normal rates.
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PMID:Antithrombin III deficiency: decreased synthesis of a biochemically normal molecule. 708 48

Heparin-Sepharose chromatography of rabbit plasma or serum yields two fractions of free antithrombin III (AT). The first of these elutes at a lower salt concentration, represents about 90% of the AT in plasma, and has an approximately 2000 dalton higher molecular weight by SDS polyacrylamide electrophoresis. An antibody to the lower affinity species reacts with the second form. The higher affinity AT is not formed from the lower affinity type during blood coagulation as demonstrated by approximately equal levels in plasma and serum, and lack of conversion of 125I-labelled lower affinity AT to the higher affinity form during blood coagulation. Heparin cofactor and progressive AT activities of the two forms are essentially identical when assayed by chromogenic substrates. The forms are separable by crossed immunoelectrophoresis.
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PMID:Isolation and partial characterization of two distinct types of antithrombin III from rabbit. 712 12

Isoelectric focusing (IEF) of the apoproteins of triglyceride rich human serum lipoproteins gives rise to the separation of some 15-20 protein bands. Three of these bands have been isolated in pure form and were characterized as isoelectric species of beta 2-glycoprotein-I (beta 2G-I). To compare the amino acid composition of these polymorphic forms with a representative specimen of beta 2G-I from total serum it was also necessary to apply a novel isolation procedure using Rivanol, perchloric acid and Heparin-Sepharose affinity chromatography. With the possible exception of the Pro content, the three isoforms were chemically and immunochemically identical. The isoelectric points of the polymorphic forms were 5.75, 6.0 and 6.2. Their molecular weight was identical by SDS polyacryl amide gel electrophoresis (54 000 D).
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PMID:Studies on the composition of the protein part of triglyceride rich lipoproteins of human serum: isolation of polymorphic forms of beta 2-glycoprotein-I. 731 80

We have isolated and characterized an antigen from normal human brain called p80, so called because it migrated with an M(r) of 80 kDa on SDS PAGE. The M(r) of 80 kDa consists of a protein of about 55-60 kDa and carbohydrate (20-25 kDa). The carbohydrate is almost entirely of the N-linked type, although a small amount of O-linked carbohydrate was detected. Cross-reactivity with monoclonal antibodies A3D8 and A1G3 showed that p80 could therefore be considered an isoform of the CD44 adhesion molecules. In addition, specific binding to hyaluronate which was not competed for by proteoglycan demonstrated that it involved different sites than the proteoglycan binding sites. We also observed that fucoidan and dextran sulphate increased the binding by 200-250% while chondroitin sulphate C also increased the binding but to a lesser extent. Heparin, heparan sulphate and chondroitin sulphates A and B did not have such an effect. The binding of p80 to hyaluronate was pH dependent with a maximum at pH 6.4. We concluded that p80 was an astrocyte specific adhesion molecule.
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PMID:A glycoprotein expressed by human fibrous astrocytes is a hyaluronate-binding protein and a member of the CD44 family. 752 50

HIV-1 reverse transcriptase from the HIV-1 strain WMF 1.13 was expressed in Escherichia coli JM 105 using a pKK233-2 vector. The bacteria were cultivated in a 20-l fermentor with 14-l net volume using M9ZB medium containing bactotryptone and yeast extract. After induction of reverse transcriptase (RT) expression by addition of isopropyl-beta-D-thiogalactopyranoside the enzyme concentration was monitored. Both soluble and inclusion-body deposited RT were detected by Western blots. Inclusion-body formation was confirmed by transmission electron microscopy. Further purification of soluble and insoluble RT was investigated. After cell desintegration by enzymatic treatment combined with osmotic shock and centrifugation, the supernatant was desalted by size-exclusion chromatography and further purified by DEAE-Sepharose FF, AF-Heparin Toyopearl 650 M and Fractogel EMD TMAE 650 (S). The results of the purification steps were monitored by SDS-PAGE with silver staining, non-radioactive RT assay and protein determination with Coomassie Blue. The sediment was extracted with 6 M GuHCl and after clarification and conventional refolding, treated in the same manner as soluble RT. This method is well suited for studying fermentation conditions as well as purification conditions. The RT is expressed in approximately equal amounts as soluble and insoluble enzyme.
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PMID:Laboratory-scale production and purification of recombinant HIV-1 reverse transcriptase. 753 52

The heart hypertrophies in response to certain forms of increased mechanical load, but it is not understood how, at the molecular level, the mechanical stimulus of increased load is transduced into a cell growth response. One possibility is that mechanical stress provokes the release of myocyte-derived autocrine growth factors. Two such candidate growth factors, acidic and basic fibroblast growth factor (aFGF and bFGF, respectively), are released via mechanically induced disruptions of the cell plasma membrane. In the present study, we demonstrate that transient, survivable disruption (wounding) of the cardiac myocyte plasma membrane is a constitutive event in vivo. Frozen sections of normal rat heart were immunostained to reveal the distribution of the wound event marker, serum albumin. Quantitative image analysis of these sections indicated that an average of 25% of the myocytes contained cytosolic serum albumin; ie, this proportion had suffered a plasma membrane wound. Wounding frequency increased approximately threefold after beta-adrenergic stimulation of heart rate and force of contraction. Heparin-Sepharose chromatography, enzyme-linked immunosorbent assay, growth assay coupled with antibody neutralization, and two-dimensional SDS-PAGE followed by immunoblotting were used to demonstrate that both aFGF and bFGF were released from an ex vivo beating rat heart. Importantly, beta-adrenergic stimulation of heart rate and force of contraction increased FGF release. Cell wounding is a fundamental but previously unrecognized aspect of the biology of the cardiac myocyte. We propose that contraction-induced cardiac myocyte wounding releases aFGF and bFGF, which then may act as autocrine growth-promoting stimuli.
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PMID:Contraction-induced cell wounding and release of fibroblast growth factor in heart. 753 17


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