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Human neutrophil plasma membrane. Specific labelling, topological distribution of proteins and surface antigen detection

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Abstract

We describe here the major protein components of a highly purified human neutrophil plasma membrane fraction analyzed by uni- and two-dimensional gel electrophoresis, as well as their glycoprotein nature as determined by PAS staining, [125I]-Con A binding and [3H]-sodium borohydride labelling. A polypeptide of about 150kDa appeared as the main Con A binding protein. The topology of the polypeptides has also been determined by protein labelling from the outside of the cell surface by lactoperoxidase catalyzed iodination and from within the bilayer by using the hydrophobic reagent [1251]-iodonaphtylazide. The antigenic features of some cell surface polypeptides have also been determined by the use of monoclonal antibodies. In this context we have detected by immunoprecipitation in human neutrophils the antigen MAC 120, previously found in monocytes and putatively associated with antigen presenting function.

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Abbreviations

Con A:

Concanavalin A

INA:

Iodonaphtylazide

mAB:

monoclonal Antibody

PAS:

Periodic Acid Schiff reaction

PBS:

Phosphate Buffered Saline

PMSF:

Phenylmethylsulfonil Fluoride

PPO:

2,5-Diphenyloxazole

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  1. Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Cientificas, Madrid, Spain

    Pedro Lacal, Faustino Mollinedo & Vicente Larraga

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  1. Pedro Lacal
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  2. Faustino Mollinedo
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  3. Vicente Larraga
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Lacal, P., Mollinedo, F. & Larraga, V. Human neutrophil plasma membrane. Specific labelling, topological distribution of proteins and surface antigen detection. Mol Cell Biochem 77, 161–171 (1987). https://doi.org/10.1007/BF00221925

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  • DOI: https://doi.org/10.1007/BF00221925

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