Two-dimensional analysis of human lymphocyte proteins II. Regulation of lymphocyte proteins by a molecule present in normal human urine.

1982 ◽  
Vol 28 (4) ◽  
pp. 1074-1083 ◽  
Author(s):  
K E Willard
Keyword(s):  
Human Urine ◽  
Degradation Products ◽  
Human Lymphocytes ◽  
Cytoskeletal Proteins ◽  
Relative Molecular Mass ◽  
Effector Protein ◽  
Two Dimensional ◽  
Effector Molecule ◽  
Normal Human ◽  
Peptide Maps

Abstract Lymphocytes cultured with concentrated human urine develop many alterations (Clin. Chem. 27:1327, 1981). New proteins appear ("Urocon" proteins), while others disappear ("Urocof" proteins) from the two-dimensional gel pattern. This paper characterizes a human urinary leukocyte effector protein ("HULEP") that is responsible for the appearance of the Urocon:11-15 proteins in human lymphocytes. The effector molecule, HULEP-1, is a protein with a relative molecular mass in the range of 29000 to 43000. Urocon:11-15 production, cytoskeletal changes in the lymphocyte, and the presence of added HULEP-1 in the cultures are all directly related. This molecule appears to alter the structure of the lymphocyte by degrading actin and possible other lymphocyte cytoskeletal proteins. Peptide maps of Urocon:11-15 and beta and gamma actin provide evidence that Urocon:11-15 proteins are closely related to nonmuscle actin, and may be degradation products of this major cytoskeletal protein.

Two-dimensional analysis of human lymphocyte proteins: I. An assay for lymphocyte effectors.

1981 ◽  
Vol 27 (8) ◽  
pp. 1327-1334 ◽  
Author(s):  
K E Willard ◽  
N G Anderson
Keyword(s):  
Transcriptional Control ◽  
Molecular Level ◽  
Human Lymphocytes ◽  
Biologically Active ◽  
Two Dimensional ◽  
Urinary Proteins ◽  
Active Protein ◽  
Two Dimensional Gel Electrophoresis ◽  
Wide Range ◽  
Normal Human

Abstract We describe an assay for lymphocyte effectors that is capable of establishing the existence of regulators of lymphocyte gene expression (including post-transcriptional control and protein processing) and has the ability to characterize the response at the molecular level. The hypothesis that circulating effectors substances excreted through the kidney can be actively present in human urine was tested with this assay. Thus, biologically active protein molecules in urine were detected at concentrations of less than 1 mg/L and over a wide range of dilutions. Activities were detected and quantitated by culturing human lymphocytes with human urinary proteins in the presence of [35S]methionine and subsequently analyzing the labeled lymphocyte proteins by two-dimensional gel electrophoresis. Thus, protein analysis by two-dimensional gels was used to indirectly detect changes produced in cultured lymphocytes after exposure to regulatory molecules. Proteins or sets of lymphocyte proteins appeared or disappeared after exposure to normal or pathological human urinary proteins. Normal human urinary proteins triggered the appearance of sets of proteins referred to by number as the "Urocon" proteins and suppressed the synthesis of protein sets referred to as "Urocof" proteins. In addition to the normal alterations described, urinary proteins from individuals with influenza or acute leukemia and after renal transplantation were capable of inducing unique alterations in lymphocyte patterns.

Excretion of X-prolyl dipeptidyl-aminopeptidase in human urine as determined with a new fluorogenic substrate.

1978 ◽  
Vol 24 (7) ◽  
pp. 1163-1166 ◽  
Author(s):  
T Kato ◽  
T Hama ◽  
K Kojima ◽  
T Nagatsu ◽  
S Sakakibara
Keyword(s):  
Enzyme Activity ◽  
Human Urine ◽  
Aminopeptidase Activity ◽  
Main Peak ◽  
Relative Molecular Mass ◽  
Fluorogenic Substrate ◽  
Km Value ◽  
Normal Human ◽  
Dipeptidyl Aminopeptidase ◽  
Normal Adults

Abstract X-Prolyl dipeptidyl-aminopeptidase activity was found in human urine by a sensitive fluorescence assay in which a new fluorogenic substrate, 7-glycylproline-4-methylcoumarinamide, is used. The Km value was 2.9 X 10(-4) mol/liter, and the optimum pH was 8.7 in glycine-NaOH buffer. The enzyme activity was stable at 4 degrees C for at least five days. On Sephadex G-200 column chromatography, normal human urine showed a main peak with an approximate relative molecular mass of 400 000. The procedure is simple, rapid, and accurate. The enzyme activity in urine of normal adults was: 4.30 +/- 0.13 (SE) (range, approximately 1.84-8.96) micronmol/min per gram of creatinine, and 2.16 +/- 0.09 (SE) (range, approximately 0.38-6.98) micronmol/min per liter of urine.

scholarly journals Proteins of human urine. III. Identification and two-dimensional electrophoretic map positions of some major urinary proteins.

1982 ◽  
Vol 28 (4) ◽  
pp. 941-948 ◽  
Author(s):  
J J Edwards ◽  
S L Tollaksen ◽  
N G Anderson
Keyword(s):  
Molecular Mass ◽  
Human Urine ◽  
Retinol Binding Protein ◽  
Immunological Methods ◽  
Relative Molecular Mass ◽  
Two Dimensional ◽  
Ph Gradients ◽  
Major Urinary Proteins ◽  
Wide Range ◽  
Beta 2 Microglobulin

Abstract We mapped the proteins of human urine by high-resolution two-dimensional electrophoresis, utilizing the ISO-DALT system. Wide-range pH gradients and narrow-range acid gradients were both used in the first-dimension separations. The patterns revealed proteins ranging in relative molecular mass from 10 000 to 90 000. Proteins identified in the map included transferrin, albumin, hemopexin, alpha 2-HS glycoprotein, alpha 1-antitrypsin. Gc globulin, alpha 1-acid glycoprotein, Zn alpha 2-glycoprotein, retinol binding protein, beta 2-microglobulin, the immunoglobulin light chains, and MAUP (most acid urinary protein). The use and utility of internal-charge and molecular-mass standards are described. We used electrophoretic transfer of proteins to nitrocellulose sheets and subsequent detection by immunological methods to identify some proteins.

A SIMPLE METHOD FOR THE DETECTION OF URINARY UNCONJUGATED CORTISOL BY PAPER CHROMATOGRAPHY

1964 ◽  
Vol 47 (3) ◽  
pp. 466-468 ◽  
Author(s):  
S. B. Pal
Keyword(s):  
Paper Chromatography ◽  
Human Urine ◽  
Specific Method ◽  
Simple Method ◽  
Normal Human

ABSTRACT Unconjugated corticosteroids are extracted from normal human urine and the urine of patients with rheumatic disorders treated with synthetic corticosteroids and corticotrophin. A simple and specific method using paper chromatography has been developed to detect the unconjugated cortisol in urine.

scholarly journals A monoclonal antibody to a human neutrophil-specific plasma membrane antigen. Effect of the antibody on the C3bi-mediated adherence by neutrophils and expression of the antigen during myelopoiesis.

1988 ◽  
Vol 167 (2) ◽  
pp. 421-439 ◽  
Author(s):  
B Pytowski ◽  
T G Easton ◽  
J E Valinsky ◽  
T Calderon ◽  
T Sun ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Bone Marrow Cells ◽  
Human Lymphocytes ◽  
Leukemic Cell ◽  
Human Neutrophils ◽  
Binding Studies ◽  
Granulocytic Differentiation ◽  
Average Molecular Mass ◽  
Cytochemical Analysis ◽  
Normal Human

We have used mice selectively tolerized to antigens of human lymphocytes by treatment with cyclophosphamide to raise an mAb, BH2-C6, that reacts with a plasma membrane antigen specific for human neutrophils. This specificity is demonstrated by indirect immunofluorescence microscopy, cytochemical analysis of fluorescence-positive and -negative cell populations separated by flow cytometry, and by the selective, complement-mediated killing of mAb BH2-C6-treated neutrophils. Additional evidence for the neutrophil specificity of mAb BH2-C6 is shown by immunoelectron microscopy, which demonstrates a lack of reactivity with human eosinophils. Immunoblotting of SDS-PAGE-separated proteins of polymorphonuclear leukocytes with 125I-labeled BH2-C6 identifies protein with an average molecular mass of 157 kD. Binding studies show that, at saturation, neutrophils bind 214,000 molecules of 125I-BH2-C6 per cell. Addition of mAb BH2-C6 to neutrophils significantly reduces the number of C3bi-opsonized sheep erythrocytes (EIgMC3bi) bound by these cells. This reduction is partly reversed by the presence of soybean trypsin inhibitor (SBTI), indicating that at least one part of this inhibition is due to BH2-C6-stimulated secretion of a serine protease that may affect ligand binding. Cytochemical analysis of normal human bone marrow cells sorted by cytofluorimetry identifies the promyelocyte as the precursor cell that first expresses BH2-Ag on the plasma membrane. Using the leukemic cell line HL-60, we demonstrate that only inducers of granulocytic differentiation, cis-retinoic acid, and dimethyloxazolidine stimulate the expression of BH2-Ag. These results show that the expression of BH2-Ag during myelomonocytic differentiation is a property uniquely possessed by cells committed to the neutrophilic lineage.

Application of HPLC-ICP-MS to speciation of cisplatin and its degradation products in water containing different chloride concentrations and in human urine

2003 ◽  
Vol 18 (11) ◽  
pp. 1391-1395 ◽  
Author(s):  
S. Hann ◽  
G. Koellensperger ◽  
Zs. Stefánka ◽  
G. Stingeder ◽  
M. Fürhacker ◽  
...  
Keyword(s):  
Human Urine ◽  
Degradation Products ◽  
Icp Ms ◽  
Chloride Concentrations
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