scholarly journals Intact rough‐ and smooth‐form lipopolysaccharides fromEscherichia coliseparated by preparative gel electrophoresis exhibit differential biologic activity in human macrophages

FEBS Journal ◽  
2013 ◽  
Vol 280 (4) ◽  
pp. 1095-1111 ◽  
Author(s):  
Elder Pupo ◽  
Buko Lindner ◽  
Helmut Brade ◽  
Andra B. Schromm
Keyword(s):  
Gel Electrophoresis ◽  
Smooth Form ◽  
Human Macrophages ◽  
Biologic Activity

scholarly journals Fibronectin is produced by human macrophages.

1980 ◽  
Vol 151 (3) ◽  
pp. 602-613 ◽  
Author(s):  
K Alitalo ◽  
T Hovi ◽  
A Vaheri
Keyword(s):  
Gel Electrophoresis ◽  
Morphological Differentiation ◽  
Mononuclear Leukocytes ◽  
No Production ◽  
Metabolic Labeling ◽  
Human Macrophages ◽  
Human Fibronectin ◽  
Enriched Cultures ◽  
Promote Cell Adhesion

Monocyte-enriched cultures were prepared from human blood mononuclear leukocytes by adherence to growth substratum. Synthesis and secretion of fibronectin was detected in these cultures concomitantly with morphological differentiation, starting on day 3--5. Production of fibronectin by macrophages was documented by metabolic labeling followed by immunoprecipitation and gel electrophoresis, radioimmunoassay specific for human fibronectin, and by indirect immunofluorescence. Fibronectin was detected mainly intracellularly but was also detected pericellularly only in minute amounts. No production of collagenous proteins was seen in these cultures. Macrophage fibronectin might act in vivo as a nonspecific opsonin and promote cell adhesion during macrophage migration in tissues.

scholarly journals The inhibition of high and low molecular weight urokinase in plasma

Blood ◽  
1980 ◽  
Vol 55 (3) ◽  
pp. 430-436
Author(s):  
G Murano ◽  
D Aronson ◽  
L Williams ◽  
L Brown
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Antithrombin Iii ◽  
Polyacrylamide Gel Electrophoresis ◽  
Low Molecular Weight ◽  
Biologic Activity ◽  
At Iii ◽  
Neutralization Process ◽  
Alpha 1 Antitrypsin ◽  
Formation Of Complexes

The rates of inhibition of high molecular weight (HMW) and low molecular weight (LMW) urokinase (UK) incubated in plasma or with purified antithrombin III (AT-III) were compared. Using a fibrinolytic assay system to determine residual biologic activity, polyacrylamide gel electrophoresis to demonstrate the formation of complexes, and selective immunoprecipitation techniques to identify the plasma inhibitors participating in the neutralization process, it was established that: (A) HMW-UK is inhibited more rapidly than LMW-UK, both in plasma and with purified AT-III; (B) heparin (3--10 U/ml accelerates the neutralization process in both systems, but only slightly; and (C) in plasma, several inhibitors, alpha 2-macroglobulin, alpha 1-antitrypsin, and antithrombin III, neutralize the activity of HMW-UK and LMW-UK.

scholarly journals The inhibition of high and low molecular weight urokinase in plasma

Blood ◽  
1980 ◽  
Vol 55 (3) ◽  
pp. 430-436 ◽  
Author(s):  
G Murano ◽  
D Aronson ◽  
L Williams ◽  
L Brown
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Antithrombin Iii ◽  
Polyacrylamide Gel Electrophoresis ◽  
Low Molecular Weight ◽  
Biologic Activity ◽  
At Iii ◽  
Neutralization Process ◽  
Alpha 1 Antitrypsin ◽  
Formation Of Complexes

Abstract The rates of inhibition of high molecular weight (HMW) and low molecular weight (LMW) urokinase (UK) incubated in plasma or with purified antithrombin III (AT-III) were compared. Using a fibrinolytic assay system to determine residual biologic activity, polyacrylamide gel electrophoresis to demonstrate the formation of complexes, and selective immunoprecipitation techniques to identify the plasma inhibitors participating in the neutralization process, it was established that: (A) HMW-UK is inhibited more rapidly than LMW-UK, both in plasma and with purified AT-III; (B) heparin (3--10 U/ml accelerates the neutralization process in both systems, but only slightly; and (C) in plasma, several inhibitors, alpha 2-macroglobulin, alpha 1-antitrypsin, and antithrombin III, neutralize the activity of HMW-UK and LMW-UK.

The Interaction of Immobilized Thrombin with Human Antithrombin III

1984 ◽  
Vol 51 (01) ◽  
pp. 027-031 ◽  
Author(s):  
E Marciniak ◽  
G Gora-Maslak
Keyword(s):  
Inhibitory Activity ◽  
Gel Electrophoresis ◽  
Antithrombin Iii ◽  
Solid Support ◽  
Protein Fragment ◽  
High Affinity ◽  
Biologic Activity ◽  
Coagulant Activity ◽  
At Iii ◽  
Sepharose 4B

SummaryInteraction of human antithrombin III (AT III) with human α-thrombin coupled to Sepharose 4B was investigated. Despite markedly reduced esterolytic, amidolytic and especially coagulant activity, more than 90% of immobilized thrombin formed stable complexes with purified AT III. Presence of high affinity heparin did not facilitate the inhibition to the degree seen in reactions conducted with soluble thrombin. Instead, heparin induced proteolysis of up to 66% of the inhibitor that remained in solution. This led to the isolation of a homogeneous protein fragment which migrated in SDS-gel electrophoresis as a band of 50,000 Mr, cross-reacted with antibodies to human AT III but showed no biologic activity nor sufficient affinity for heparin. Out of the three major inhibitors capable of binding soluble thrombin in human plasma, only AT III reacted with immobilized thrombin. However, Sepharose-coupled thrombin mixed with plasma in the presence of heparin produced outstanding quantities of residual immunoreactive AT III devoid of inhibitory activity. These data suggest that presence of high affinity heparin in the environment of thrombin attached to a solid support may dramatically decrease the efficiency of enzyme inhibition.

Ultrastructural localization of specific nucleoprotein fractions

1978 ◽  
Vol 36 (2) ◽  
pp. 204-205
Author(s):  
G. L. Brown
Keyword(s):  
Gel Electrophoresis ◽  
Mouse Liver ◽  
Polyacrylamide Gel Electrophoresis ◽  
Ultrastructural Localization ◽  
Polyacrylamide Gel ◽  
Acid Extraction ◽  
Acid Solutions ◽  
Low Salt ◽  
Liver Nuclei ◽  
Phosphate Groups

Bismuth (Bi) stains nucleoproteins (NPs) by interacting with available amino and primary phosphate groups. These two staining mechanisms are distinguishable by glutaraldehyde crosslinking (Fig. 1,2).Isolated mouse liver nuclei, extracted with salt and acid solutions, fixed in either formaldehyde (form.) or gl utaraldehyde (glut.) and stained with Bi, were viewed to determine the effect of the extractions on Bi stainina. Solubilized NPs were analyzed by SDS-polyacrylamide gel electrophoresis.Extraction with 0.14 M salt does not change the Bi staining characteristics (Fig. 3). 0.34 M salt reduces nucleolar (Nu) staining but has no effect on interchromatinic (IC) staining (Fig. 4). Proteins responsible for Nu and glut.- insensitive IC staining are removed when nuclei are extracted with 0.6 M salt (Fig. 5, 6). Low salt and acid extraction prevents Bi-Nu staining but has no effect on IC staining (Fig. 7). When nuclei are extracted with 0.6 M salt followed by low salt and acid, all Bi-staining components are removed (Fig. 8).

Molecular Structure of the Outermost Cell Wall Component of Spirillum Serpens

1977 ◽  
Vol 35 ◽  
pp. 342-343
Author(s):  
Wah Chiu ◽  
David Grano
Keyword(s):  
Molecular Structure ◽  
Cell Wall ◽  
Outer Membrane ◽  
Gel Electrophoresis ◽  
Electron Microscopic Examination ◽  
Electron Microscopic ◽  
Cell Wall Component ◽  
Protein Components ◽  
Exposure Technique ◽  
Structural Aspects

The periodic structure external to the outer membrane of Spirillum serpens VHA has been isolated by similar procedures to those used by Buckmire and Murray (1). From SDS gel electrophoresis, we have found that the isolated fragments contain several protein components, and that the crystalline structure is composed of a glycoprotein component with a molecular weight of ∽ 140,000 daltons (2). Under an electron microscopic examination, we have visualized the hexagonally-packed glycoprotein subunits, as well as the bilayer profile of the outer membrane. In this paper, we will discuss some structural aspects of the crystalline glycoproteins, based on computer-reconstructed images of the external cell wall fragments.The specimens were prepared for electron microscopy in two ways: negatively stained with 1% PTA, and maintained in a frozen-hydrated state (3). The micrographs were taken with a JEM-100B electron microscope with a field emission gun. The minimum exposure technique was essential for imaging the frozen- hydrated specimens.

Interrelationship of the cellular distribution and secretion of regular structure (RS) protein in Bacillus licheniformis nm 105

1992 ◽  
Vol 50 (1) ◽  
pp. 712-713
Author(s):  
Xiaorong Zhu ◽  
Richard McVeigh ◽  
Bijan K. Ghosh
Keyword(s):  
Alkaline Phosphatase ◽  
Bacillus Licheniformis ◽  
Self Assembly ◽  
Gel Electrophoresis ◽  
Culture Supernatant ◽  
Regular Structure ◽  
The Self ◽  
Cellular Distribution ◽  
Structural Protein ◽  
Laboratory Cultures

A mutant of Bacillus licheniformis 749/C, NM 105 exhibits some notable properties, e.g., arrest of alkaline phosphatase secretion and overexpression and hypersecretion of RS protein. Although RS is known to be widely distributed in many microbes, it is rarely found, with a few exceptions, in laboratory cultures of microorganisms. RS protein is a structural protein and has the unusual properties to form aggregate. This characteristic may have been responsible for the self assembly of RS into regular tetragonal structures. Another uncommon characteristic of RS is that enhanced synthesis and secretion which occurs when the cells cease to grow. Assembled RS protein with a tetragonal structure is not seen inside cells at any stage of cell growth including cells in the stationary phase of growth. Gel electrophoresis of the culture supernatant shows a very large amount of RS protein in the stationary culture of the B. licheniformis. It seems, Therefore, that the RS protein is cotranslationally secreted and self assembled on the envelope surface.

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