Determination of the Molecular Weights of Membrane Proteins and Polypeptides

Fish, Wayne W.

doi:10.1007/978-1-4684-2907-7_4

Cited by 5 publications

(4 citation statements)

References 115 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The inaccuracy in determining the molecular weight of a nonreduced protein by SDS-PAGE is well known (9). However, the apparent doubling of the molecular weight of the HN glycoprotein in the absence of reducing agent has been noted for other paramyxoviruses, suggesting that the native protein on the surface of these viruses is a dimer linked by disulfide bonds (10,13,25,33).…”

Section: Resultsmentioning

confidence: 99%

Differentiation of mumps virus strains with monoclonal antibody to the HN glycoprotein

Server¹,

Merz²,

Waxham³

et al. 1982

Infect Immun

View full text Add to dashboard Cite

A hybridoma cell line secreting antibody of the immunoglobulin G3 isotype with kappa light chains and with activity against the HN glycoprotein of the Kilham strain of mumps virus was established. The antibody exhibited structural homogeneity in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and had a microheterogeneous isoelectric spectrum characteristic of an antibody of monoclonal origin. The specificity of the monoclonal antibody, shown by immunoprecipitation performed with radiolabeled virus and infected cell lysates, was for the larger mumps virus glycoprotein. In functional assays the antibody inhibited the hemagglutinating and neuraminidase activities and neutralized the infectivity of the homologous Kilham strain of virus and clearly differentiated this strain from two heterologous strains, Enders and O'Take. The antibody was markedly less effective with the O'Take strain than with either the Kilham or Enders strain in inhibiting both hemagglutination and neuraminidase activity against the macromolecular substrate fetuin. The inhibition of the neuraminidase activity of the Kilham strain was independent of substrate size, the antibody inhibiting the hydrolysis of both fetuin and the trisaccharide neuraminlactose. By contrast, the antibody did not inhibit the hydrolysis of neuraminlactose by the two heterologous mumps strains. These results provide the first demonstration of antigenic differences between mumps virus strains and highlight the utility of monoclonal antibody in analyzing the structural basis underlying functional activities of the HN glycoproteins.

show abstract

Section: Resultsmentioning

confidence: 99%

Differentiation of mumps virus strains with monoclonal antibody to the HN glycoprotein

Server¹,

Merz²,

Waxham³

et al. 1982

Infect Immun

View full text Add to dashboard Cite

show abstract

“…This aberrantly rapid migration would lead to underestimates of the true molecular weights of the arylsulfatase oligomers and is contrary to the usual overestimation of molecular weights of glycoproteins on SDS-polyacrylamide gel electrophoresis (24). Aberrantly rapid mobility (25) has been recorded previously with proteins binding quantities of SDS in excess of 1.4 g SDS/g of protein (26), for ferridoxins due to the inherent charge of acidic residues (27), and for a few proteins without explanation (28) as is the situation with eosinophil arylsulfatase B. Apparent molecular weights by SDS-PAGE, as in Fig.…”

Section: Purification Of Human Eosinophil Arylsulfatase Bmentioning

confidence: 94%

Human eosinophil arylsulfatase B. Structure and activity of the purified tetrameric lysosomal hydrolase.

Weller¹,

Austen²

1983

J. Clin. Invest.

View full text Add to dashboard Cite

A B S T R A C T Arylsulfatase B from human eosinophils was purified free of contaminating proteins by gel filtration and sequential affinity chromatography on Affi-Gel Blue and zinc chelate Sepharose. 50 Ag of the purified enzyme presented as a single stained band on alkaline disc gel electrophoresis. In both goats and rabbits, the purified enzyme elicited monospecific antisera that yielded single precipitation arcs on Ouchterlony analysis with a human eosinophil extract and the purified enzyme; the immunoprecipitation lines fused in a pattern of identity, providing immunochemical evidence for the homogeneity of the purified enzyme. On sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, a dominant lower molecular weight protein and three other bands with molecular weights approximately two, three, and four times that of the major protein band were resolved. The prominence of the less rapidly migrating protein bands increased relative to the major band if the enzyme was maintained under acidic conditions or was reacted with the crosslinking agent dimethyl suberimidate under alkaline conditions before SDS-polyacrylamide gel electrophoresis, supporting the conclusion that the enzyme consists of four subunits. Two stained bands were present on acid disc gel electrophoresis; they were composed of oligomeric forms of enzyme on analysis by SDSpolyacrylamide gel electrophoresis in a second dimension. A minimum molecular weight of 70,190 was de-

show abstract

“…The most relevant details will be recapitulated. The preparation of the sample is adapted to the factors relevant to the formation of SDS protein complexes (Fish, 1975).…”

Section: Sample Preparation and Sds Gel Filtrationmentioning

confidence: 99%

Relative synthesis rate of individual muscle proteins: a new approach

Schreurs¹,

Boekholt²,

Koopmanschap³

et al. 1985

NJAS

View full text Add to dashboard Cite

This paper describes the estimation of the relative synthesis rate of muscle proteins by means of a new approach. This approach is an obvious alternative for isolation procedures, with usually low protein recovery. Loss of protein material is avoided by taking advantage of separating the muscle proteins in a single step by SDS gel fil tration. As a consequence an overall impression of the relative synthesis rate of the main muscle proteins can be visualized. This can be useful in comparative studies describing the influence of physiological parameters on the synthesis of individual muscle proteins.

show abstract

Determination of the Molecular Weights of Membrane Proteins and Polypeptides

Cited by 5 publications

References 115 publications

Differentiation of mumps virus strains with monoclonal antibody to the HN glycoprotein

Differentiation of mumps virus strains with monoclonal antibody to the HN glycoprotein

Human eosinophil arylsulfatase B. Structure and activity of the purified tetrameric lysosomal hydrolase.

Relative synthesis rate of individual muscle proteins: a new approach

Contact Info

Product

Resources

About