Distribution and metabolism of protein antigens in rat liver*1

Perlmann, Peter; Hultin, T.; D’Amelio, V.; Morgan, Winfield S.

doi:10.1016/0014-4827(59)90249-6

Cited by 17 publications

(10 citation statements)

References 21 publications

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“…THE ANTISERA: As prcviously described, injection into rabbits of rat liver microsomes or cell sap leads to the production of antisera possessing a fair degree of "fraction specificity" (1,19,21). The patterns of precipitates obtained with the antimicrosomal and the anti-cell sap scra of the present study arc seen below in Figs.…”

Section: E S C R I P T I O N Of T H E R E a G E N T Ssupporting

confidence: 58%

“…The insoluble portions of the microsomes were separated by centrifugation at 105,000 g for 2 hours. The soluble portion, in the following called Lubrol extract, was assumed to contain proteins solubilized from the microsomal membranes as well as possible contents of the mierosomal vesicles (18,21).…”

Section: Methodsmentioning

confidence: 99%

“…None of the precipitates seen in these figures is due to reactions between scrum protcins and their antibodies, since the latter had been removed by previous absorption. For further details see references (1,19,21). In addition to these two antiscra, one against the p H 5 precipitablc fraction of the cell sap was used.…”

Section: E S C R I P T I O N Of T H E R E a G E N T Smentioning

confidence: 99%

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Autoradiographic Analysis on Agar Plates of Antigens From Sub Cellular Fractions of Rat Liver Slices

Morgan

Perlmann

Hultin

1961

The Journal of Cell Biology

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Slices of rat livers were incubated with 14C amino acids, homogenized, and subjected to differential centrifugation. The microsomes were further extracted with the non-ionic detergent Lubrol W and with EDTA. These extracts and the microsome free "cell sap," freed from the pH 5 precipitable fraction, were subsequently reacted with antisera using agar diffusion techniques. The antisera employed were obtained from rabbits injected with different subcellular fractions of rat liver or with rat serum proteins. When the agar diffusion plates were autoradiographed it was found that some of the precipitates were radioactive while others were not. Control experiments indicated that this labeling was due to the specific incorporation of 14C amino acids into various rat liver antigens during incubation of the slices rather than to a non-specific adsorption of radioactive material to the immunological precipitates. When the slices were incubated with the isotope for up to 30 minutes, the serum proteins which could be extracted from the microsomes with the detergent were strongly labeled, as were a number of additional microsomal antigens of unknown significance. In contrast, the serum proteins present in the cell sap were only weakly labeled. Most of the typical cell sap proteins, both those precipitable and those soluble at pH 5, seemed to remain unlabeled. No consistently reproducible results were obtained with the EDTA extracts of the ribosomal residues remaining after extraction of the microsomes with the detergent. Incubation of the liver slices for longer periods (up to 120 minutes) led to a strong labeling of the serum proteins in the cell sap as well as to the appearance of labeling in additional cell sap proteins. The results are discussed with regard to the subcellular site of synthesis and the metabolism of the different antigens.

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Section: E S C R I P T I O N Of T H E R E a G E N T Ssupporting

confidence: 58%

Section: Methodsmentioning

confidence: 99%

Section: E S C R I P T I O N Of T H E R E a G E N T Smentioning

confidence: 99%

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Autoradiographic Analysis on Agar Plates of Antigens From Sub Cellular Fractions of Rat Liver Slices

Morgan

Perlmann

Hultin

1961

The Journal of Cell Biology

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“…It has been shown in a number of publications (23,24,(27)(28)(29) that the pattern of protein labeling in an amino acid-incorporating experiment can be studied by combining immunological reactions in agar gel with autoradiography. Most of these reports concern incorporating experiments with the 12,000 g supernatant of a rat liver homogenate or a microsomal-cell sap system.…”

Section: Discussionmentioning

confidence: 99%

“…For this purpose, advantage has been taken of a method worked out by Perlmann et al (23,24,(27)(28)(29). This technique combines immunoelectropboresis in agar gel with autoradiography.…”

Section: Introductionmentioning

confidence: 99%

Labeling With 14c Amino Acids of Albumin-Like Protein by Rat Liver Ribonucleoprotein Particles

Decken

1963

The Journal of Cell Biology

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Ribonucleoprotein particles were prepared by treatment of rat liver microsomes with detergents and high concentrations of KCI. They were active in incorporating 14C amino acids into protein when incubated with cell sap together with ATP, GTP, and a system to regenerate the triphosphates. The albumin of the incubation mixture, soluble at I05,000 g, and that of the fraction released by ultrasonication of the particles were studied by immunoclectrophoresis in agar gel. When the ribonucleoprotein particles were incubated with cell sap the immunological precipitation lines formed with antiserum to rat serum albumin were highly radioactive as tested by autoradiography. After zone electrophorcsis on cellulose acetate, two immunologically reactive albumins were obtained which differed in their electrophoretic mobility from rat serum albumin. Labeled albumin, when purified on DEAE-cellulose columns, retained its radioactivity as tested by autoradiography following immunoelcctrophoresis. On cellulose acetate this purified albumin showed an electrophoretic mobility higher than that of rat serum albumin.

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Cytoplasmic antigens of the liver

Kashkin¹

1967

Bull Exp Biol Med

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Distribution and metabolism of protein antigens in rat liver*1

Cited by 17 publications

References 21 publications

Autoradiographic Analysis on Agar Plates of Antigens From Sub Cellular Fractions of Rat Liver Slices

Autoradiographic Analysis on Agar Plates of Antigens From Sub Cellular Fractions of Rat Liver Slices

Labeling With 14c Amino Acids of Albumin-Like Protein by Rat Liver Ribonucleoprotein Particles

Cytoplasmic antigens of the liver

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