The Isolation and Characterization of the Soluble and Membrane-Bound Porcine Cytochrome b5cDNAs

1997 ◽  
Vol 240 (1) ◽  
pp. 80-83 ◽  
Author(s):  
Petra K. Vandermark ◽  
Alan W. Steggles
Keyword(s):  
Isolation And Characterization ◽  
Membrane Bound

scholarly journals Isolation and characterization of a membrane protein from rat erythrocytes which inhibits lysis by the membrane attack complex of rat complement

1992 ◽  
Vol 284 (1) ◽  
pp. 169-176 ◽  
Author(s):  
T R Hughes ◽  
S J Piddlesden ◽  
J D Williams ◽  
R A Harrison ◽  
B P Morgan
Keyword(s):  
Affinity Purification ◽  
Membrane Attack Complex ◽  
Erythrocyte Membranes ◽  
Dependent Manner ◽  
Inhibitory Protein ◽  
Rat Erythrocytes ◽  
Butanol Extract ◽  
Isolation And Characterization ◽  
Membrane Bound

The membrane attack complex (MAC) of complement in humans is regulated by several membrane-bound proteins; however, no such proteins have so far been described in other species. Here we report the isolation and characterization of a rat erythrocyte membrane glycoprotein of molecular mass 21 kDa which inserts into cell membranes and is a potent inhibitor of the rat MAC. This protein, here called rat inhibitory protein (RIP), was first partially purified by column chromatography from a butanol extract of rat erythrocyte membranes. Monoclonal antibodies (Mabs) were raised against RIP and used for its affinity purification. Affinity-purified RIP was shown to inhibit in a dose-dependent manner the cobra venom factor (CVF)-mediated ‘reactive’ lysis of guinea pig erythrocytes by rat complement. Conversely, the anti-RIP MAbs 6D1 and TH9 were shown to markedly enhance the CVF-mediated lysis of rat erythrocytes by rat complement. RIP acted late in the assembly of the MAC (at or after the C5b-8 stage) and was releasable from the membranes of rat erythrocytes by phosphatidylinositol-specific phospholipase C. These features, together with its size, deglycosylation pattern and N-terminal amino acid sequence, lead us to conclude that RIP is the rat homologue of the human MAC-inhibitory protein CD59 antigen.

scholarly journals Isolation and Characterization of Membrane-Bound Arylamidases from Human Placenta and Kidney

1980 ◽  
Vol 104 (1) ◽  
pp. 155-165 ◽  
Author(s):  
Kunio HIWADA ◽  
Taketoshi ITO ◽  
Masako YOKOYAMA ◽  
Tatsuo KOKUBU
Keyword(s):  
Human Placenta ◽  
Isolation And Characterization ◽  
Membrane Bound

Biosynthesis of Spin-Labelled Lipids and Their Use in the Study of Biological Membranes: Isolation and Characterization of Membrane-Bound Spin-Labelled sn-3-Phosphatidic Acid-2-3H Prepared by Enzymatic Incorporation of Spin-Labelled Stearic Acid into sn-Glycero-3-phosphate-2-3H

1974 ◽  
Vol 52 (10) ◽  
pp. 884-893 ◽  
Author(s):  
N. Z. Stanacev ◽  
L. Stuhne-Sekalec ◽  
S. Schreier-Muccillo ◽  
I. C. P. Smith
Keyword(s):  
Phosphatidic Acid ◽  
Spin Label ◽  
Liver Microsomes ◽  
Biological Membranes ◽  
General Application ◽  
Isolation And Characterization ◽  
Spin Resonance ◽  
Membrane Bound ◽  
Guinea Pig Liver Microsomes

Spin-labelled 12-doxylstearic acid was covalently incorporated into sn-glycero-3-phosphate-2-3H yielding sn-3-phosphatidic acid-2-3H, a key intermediate in phospholipid biosynthesis, in a reaction catalyzed by isolated guinea-pig liver microsomes. It was found that at least 17.1% of the obtained sn-3-phosphatidic acid-2-3H contained spin label. Degradation of spin-labelled sn-3-phosphatidic acid-2-3H with the phospholipase A from Crotalus adamanteus yielded both lyso-sn-3-phosphatidic acid-2-3H and free fatty acids which were paramagnetic, establishing that the incorporated 12-doxylstearic acid occupies both sn-1- and sn-2-positions of the biosynthesized sn-3-phosphatidic acid-2-3H.A procedure for the preparation and isolation of biosynthetically spin-labelled radioactive sn-3-phosphatidic acid bound to the microsomal membrane was developed and the concentrations of spin-label and tritium were quantitatively determined. This procedure is believed to have a general application in the study of biological membranes. The electron spin resonance spectra and their quantitation are given and discussed for the spin-labelled sn-3-phosphatidic acid-2-3H in solution and in the membrane-bound form. Some general requirements for covalent spin labelling of biological membranes are discussed.

scholarly journals Isolation and characterization of an age-related antigen present on senescent human red blood cells

Blood ◽  
1981 ◽  
Vol 58 (2) ◽  
pp. 341-349
Author(s):  
EM Alderman ◽  
HH Fudenberg ◽  
RE Lovins
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Double Immunodiffusion ◽  
Specific Density ◽  
Human Red Blood Cells ◽  
Rbc Membrane ◽  
Isolation And Characterization ◽  
Age Related ◽  
Membrane Bound

Autologous membrane-bound IgG was isolated from a subpopulation of human red blood cells (RBC) with specific density greater than 1.110, by affinity chromatography of purified RBC membrane glycoprotein preparations using immobilized wheat germ agglutinin and immobilized anti-human immunoglobulin (Ig) as immunoabsorbents. The Ig-containing population thus obtained, when further separated by chromatography on Sephadex G-200 in the presence of chaotropic agents, yielded four peaks (Ia, Ib, II, and III). Double immunodiffusion revealed the presence of Ig in the first three peaks (IgM in peak Ia, IgA in Ib, and IgG in II) but not in peak III. Peak III was precipitated by the Ig-containing peaks (Ia, Ib, and II) in immunodiffusion assays, suggesting that the antigenic membrane determinants responsible for the binding of autologous Ig to senescent human RBC were contained in this peak (III). Peaks Ia, Ib and II precipitate purified asialoglycophorin; peak III was reactive with purified autoantibodies directed against asialoglycophorin. These results suggest that an age-related antigenic determinant(s) present on senescent human RBC is exposed by desialylation of the major sialoglycoprotein component of the RBC membrane.

Isolation and characterization of a membrane-bound proteinase from rat liver

1976 ◽  
Vol 177 (2) ◽  
pp. 355-363 ◽  
Author(s):  
Mirna Jusic ◽  
Silvia Seifert ◽  
Erich Weiss ◽  
Rainer Haas ◽  
Peter C. Heinrich
Keyword(s):  
Rat Liver ◽  
Isolation And Characterization ◽  
Membrane Bound

Isolation and characterization of an UDPG-dependent glucosyltransferase activity from Rauwolfia serpentina Benth. cell suspension cultures

1994 ◽  
Vol 72 (1) ◽  
pp. 51-55 ◽  
Author(s):  
Ralf Lutterbach ◽  
Carl Michael Ruyter ◽  
Joachim Stöckigt
Keyword(s):  
Enzyme Activity ◽  
Cell Suspension ◽  
Cell Suspension Cultures ◽  
Callus Cultures ◽  
Suspension Cultures ◽  
Rauwolfia Serpentina ◽  
Isolation And Characterization ◽  
Membrane Bound ◽  
Plant Families

From cell suspension cultures of Rauwolfia serpentina Benth. a new enzyme activity was isolated and its properties determined. The enzyme is a soluble protein and catalyzes the transfer of a glucose moiety from UDPG to a wide variety of phenolic compounds with p-nitrophenol as one of the best substrates (Km = 1.21 mM, UDPG = 0.54 mM). In contrast to the membrane-bound UDPG: vomilenine-21-OH-β-D-glucosyltransferase from Rauwolfia serpentina cells, this enzyme is not able to glucosylate indole alkaloids. The enzyme activity has been detected in 14 callus cultures belonging to 10 different plant families.

scholarly journals Isolation and Characterization of Tributyltin Resistant Mutants of Escherichia coli

1984 ◽  
Vol 39 (3-4) ◽  
pp. 293-299 ◽  
Author(s):  
Akhand P. Singh ◽  
Kiran Singh
Keyword(s):  
Escherichia Coli ◽  
Class Ii ◽  
Class I ◽  
Isolation And Characterization ◽  
Membrane Bound ◽  
Resistant Mutants ◽  
K 12 ◽  
Spontaneous Mutants ◽  
Wild Type Parent

Two classes of tributyltin (TBT) resistant, spontaneous mutants of Escherichia coli K-12 were isolated, using a cytochrome containing (W 1485) and a cytochrome deficient (SASX76) strain. In contrast to the cytochrome sufficient strain, the cytochrome deficient strain was found to be fifty times more sensitive to TBT. The class I mutants, isolated from strain W 1485, also showed crossresistance to triphenyltin (TPT). As compared to its wild type parent, the TBT-resistant mutants exhibited mucoid colony type, aberrant cell morphology and reduced uptake of TPT. Based on these results, it was suggested that the resistance of class I mutants to TBT may be associated with above mentioned alterations. The class II TBT-resistant mutants were isolated from the cytochrome deficient strain, SASX76. In comparison to class I mutants, these class II mutants were found to have TBT-resistant membrane bound adenosine triphosphatase (ATPase) which may account for their resistance to TBT

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