Reconstitution of pig lymphocyte plasma membranes from solubilized components, with particular reference to membrane-associated immunoglobulins

Chavin, Stephen I.; Holliman, Andrew

doi:10.1042/bj1520267

Cited by 8 publications

(2 citation statements)

References 16 publications

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“…SDS gel electrophoresis of the lymphocyte plasma membrane preparations reproducibly revealed major peptides of 200-, 83-, 72-, 58-, and 45-kD together with minor components of 125-, 56-, 43-, 37-, and 34-kD, in a calcium-independent manner. Due to discrepancies in molecular mass assignments between different laboratories, it is difficult to compare these results exactly with others, but the overall pattern is very similar to that seen by a number of laboratories (21,31,32). The peptides at 200 and 45 kD are almost certainly assignable to myosin heavy chain and actin, respectively (31,33).…”

Section: Discussionmentioning

confidence: 77%

Calcium-dependent association of a protein complex with the lymphocyte plasma membrane: probable identity with calmodulin-calcineurin.

Chantler¹

1985

The Journal of Cell Biology

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A protein complex is shown to participate in a calcium-dependent association with plasma membranes purified either from pig mesenteric lymph node lymphocytes or from human lymphoblastoid cell lines. Plasma membranes prepared in the presence of calcium possess this complex; those prepared in the absence of calcium (5 mM EGTA) do not. The complex associates itself with the inner cytoplasmic surface of the plasma membrane. This complex is referred to as the "acidic protein band" because of its location during migration upon alkaline-urea gel electrophoresis. The complex dissociates from the plasma membrane during electrophoresis on 8-M urea gels, irrespective of calcium levels during electrophoresis; at intermediate urea concentrations (4-6 M), the complex is not dissociated in the presence of calcium. Upon purification of the acidic protein band, SDS acrylamide gel electrophoresis, immunoblotting, and radioimmunoassay techniques suggest that the acidic protein band is composed of at least four peptides (designated 68K, 59K, 20K, 20K): two of these (68K, 20K) are immunopositive for calcineurin and one (20K) is immunopositive for calmodulin. Immunoblots of urea gels also indicate that the calcineurin heavy chain (68K) can also appear at three different locations on the urea gel. Patches and caps induced in human peripheral blood lymphocytes by fluorescein-conjugated goat anti-human IgG are not coincident with the location of calcineurin, which remains distributed throughout the cell.

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Section: Discussionmentioning

confidence: 77%

Calcium-dependent association of a protein complex with the lymphocyte plasma membrane: probable identity with calmodulin-calcineurin.

Chantler¹

1985

The Journal of Cell Biology

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“…Highly purified preparations of pig lymphocyte plasma membrane contain a major Coomassie-Blue-staining component of mol.wt. about 68000, as judged by SDS/polyacrylamide-gel electrophoresis under reducing conditions (Allan & Crumpton, 1971;Chavin & Holliman, 1975;Walsh et al, 1976). On the basis of the relative intensities of Coomassie Blue staining, this protein is present to a similar extent to actin, which has been estimated to represent 5-6% of the total membrane protein ).…”

mentioning

confidence: 99%

Albumin associated with purified pig lymphocyte plasma membrane

Barber¹,

Faulkes²,

Crumpton³

1980

Biochemical Journal

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Plasma-membrane preparations purified from pig lymphocytes contained a major polypeptide component of mol.wt. about 68 000. This component was identified as pig albumin by the following comparisons with authentic pig serum albumin: (a) co-migration when analysed by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis under reducing and non-reducing conditions; (b) identical isoelectric points; (c) similar "fingerprints" of arginine-containing tryptic peptides; (d) reactivity with anti-(pig albumin) serum. The albumin was bound tightly to the plasma membrane. Biosynthetic labelling of pig lymphocytes under a variety of conditions failed to provide evidence that albumin was synthesized by lymphocytes, suggesting that the plasma-membrane-associated albumin was of extraneous origin. Radiolabelled pig serum albumin, however, failed to bind to the plasma-membrane fraction when added before cell disruption. Although lymphocyte plasma membrane preparations from other species possessed a polypeptide of about 68 000 mol.wt., this was judged not to be albumin on the basis of electrophoretic mobility under non-reducing conditions; also, no polypeptide was precipitated by anti-albumin sera. It is concluded that pig lymphocyte plasma-membrane preparations possess albumin which, although firmly attached, was probably of extraneous origin. This association appeared not to be common to lymphocytes from other species.

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Effector Sites on Antibodies

Calvanico

TomasiJr.

1979

Immunochemistry of Proteins

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Reconstitution of pig lymphocyte plasma membranes from solubilized components, with particular reference to membrane-associated immunoglobulins

Cited by 8 publications

References 16 publications

Calcium-dependent association of a protein complex with the lymphocyte plasma membrane: probable identity with calmodulin-calcineurin.

Calcium-dependent association of a protein complex with the lymphocyte plasma membrane: probable identity with calmodulin-calcineurin.

Albumin associated with purified pig lymphocyte plasma membrane

Effector Sites on Antibodies

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