Purification, location, and immunological characterization of the iron-regulated high-molecular-weight proteins of the highly pathogenic yersiniae.

SummaryUrokinase (UK) could be purified to apparent homogeneity starting from crude urine by sequential adsorption and elution of the enzyme to gelatine-Sepharose and agmatine-Sepharose followed by gel filtration on Sephadex G-150. The purified product exhibited characteristics of the high molecular weight urokinase (HMW-UK) but did contain two distinct entities, one of which exhibited a two chain structure as reported for the HMW-UK while the other one exhibited an apparent single chain structure. The purification described is rapid and simple and results in an enzyme with probably no major alterations. Yields are high enough to obtain purified enzymes for characterization of UK from individual donors.

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INTERFERENCE BY SERUM PROTEINS WITH LH RADIOIMMUNOASSAY USING IMMUNOSORBENT

Acta Endocrinologica ◽

10.1530/acta.0.0720235 ◽

1973 ◽

Vol 72 (2) ◽

pp. 235-242 ◽

Cited By ~ 11

Author(s):

A. M. Reuter ◽

J. C. Hendrick ◽

J. Sulon ◽

P. Franchimont

Keyword(s):

Molecular Weight ◽

Human Serum ◽

High Molecular Weight ◽

Serum Proteins ◽

Void Volume ◽

High Molecular Weight Proteins ◽

Serum Fractionation ◽

Covalently Bound

ABSTRACT The percentage of LH* bound to antibodies that have been covalently bound to cellulose is diminished in the presence of LH-free human serum and sera from various species of animals. Serum fractionation studies on Sephadex G 200 show that the greatest interference comes from the proteins eluted in the void volume i. e. the high molecular weight proteins. Specifically, the gamma M globulins and the α2-macroglobulins appear to play an important role, as demonstrated by tests in which these proteins were neutralized by gamma M and α2-macroglobulin antisera.

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Faculty Opinions recommendation of Quantitative NMR studies of high molecular weight proteins: application to domain orientation and ligand binding in the 723 residue enzyme malate synthase G.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1013349.190158 ◽

2003 ◽

Author(s):

H Jane Dyson

Keyword(s):

Molecular Weight ◽

Ligand Binding ◽

High Molecular Weight ◽

Malate Synthase ◽

Nmr Studies ◽

Quantitative Nmr ◽

Domain Orientation ◽

Malate Synthase G ◽

High Molecular Weight Proteins

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Preparation and Characterization of the High Molecular Weight [3H]Hyaluronic Acid

Collection of Czechoslovak Chemical Communications ◽

10.1135/cccc19922151 ◽

1992 ◽

Vol 57 (10) ◽

pp. 2151-2156 ◽

Cited By ~ 2

Author(s):

Peter Chabreček ◽

Ladislav Šoltés ◽

Hynek Hradec ◽

Jiří Filip ◽

Eduard Orviský

Keyword(s):

Molecular Weight ◽

Hyaluronic Acid ◽

High Molecular Weight ◽

Methyl Bromide ◽

High Performance ◽

Hydrogen Atoms ◽

Isolation And Characterization ◽

Labelling Procedure ◽

Enzymatic Depolymerization

Two methods for the preparation of high molecular weight [3H]hyaluronic acid were investigated. In the first one, hydrogen atoms in the molecule were replaced by tritium. This isotopic substitution was performed in aqueous solution using Pd/CaCO3 as the catalyst. In the second method, the high molecular weight hyaluronic acid was alkylated with [3H]methyl bromide in liquid ammonia at a temperature of -33.5 °C. High-performance gel permeation chromatographic separation method was used for the isolation and characterization of the high molecular weight [3H]hyaluronic acid. Molecular weight parameters for the labelled biopolymers were Mw = 128 kDa, Mw/Mn = 1.88 (first method) and Mw = 268 kDa, Mw/Mn = 1.55 (second method). The high molecular weight [3H]hyaluronic acid having Mw = 268 kDa was degraded further by specific hyaluronidase. Products of the enzymatic depolymerization were observed to be identical for both, labelled and cold biopolymer. This finding indicates that the described labelling procedure using [3H]methyl bromide does not induce any major structural rearrangements in the molecule.

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