Polymorphism in the M sub r 32,000 Rh protein purified from Rh(D)-positive and -negative erythrocytes

Saboori, A M; Smith, B L; Agre, P

doi:10.1073/pnas.85.11.4042

Title: Polymorphism in the M sub r 32,000 Rh protein purified from Rh(D)-positive and -negative erythrocytes

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Abstract

A M{sub r} 32,000 integral membrane protein has previously been identified on erythrocytes bearing the Rh(D) antigen and is thought to contain the antigenic variations responsible for the different Rh phenotypes. To study it on a biochemical level, a simple large-scale method was developed to purify the M{sub r} 32,000 Rh protein from multiple units of Rh(D)-positive and -negative blood. Erythrocyte membrane vesicles were solubilized in NaDodSO{sub 4}, and a tracer of immunoprecipitated {sup 125}I surface-labeled Rh protein was added. The Rh protein was purified to homogeneity by hydroxylapatite chromatography followed by preparative NaDodSO{sub 4}/PAGE. Approximately 25 nmol of pure Rh protein was recovered from each unit of Rh(D)-positive and -negative blood. Rh protein purified from both Rh phenotypes appeared similar by one-dimensional NaDodSO{sub 4}/PAGE, and the N-terminal amino acid sequences for the first 20 residues were identical. Rh proteins purified from Rh(D)-positive and -negative blood were compared by two-dimensional iodopeptide mapping after {sup 125}I-labeling and {alpha}-chymotrypsin digestion. The peptide maps were very similar. These data indicate that a similar core Rh protein exists in both Rh(D)-positive and -negative erythrocytes, and the Rh proteins from erythrocytes with different Rh phenotypes contain distinct structural polymorphisms.

Authors:

Saboori, A M; Smith, B L; Agre, P ^[1]

Johns Hopkins Univ. School of Medicine, Baltimore, MD (USA)

Publication Date:: Wed Jun 01 00:00:00 EDT 1988

OSTI Identifier:: 5565401

Resource Type:: Journal Article

Journal Name:: Proceedings of the National Academy of Sciences of the United States of America; (USA)

Additional Journal Information:: Journal Volume: 85:11; Journal ID: ISSN 0027-8424

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES; ANTIGENS; MOLECULAR STRUCTURE; PROTEINS; PURIFICATION; BLOOD GROUPS; CELL MEMBRANES; ELECTROPHORESIS; ERYTHROCYTES; IODINE 125; PHENOTYPE; BETA DECAY RADIOISOTOPES; BIOLOGICAL MATERIALS; BLOOD; BLOOD CELLS; BODY FLUIDS; CELL CONSTITUENTS; DAYS LIVING RADIOISOTOPES; ELECTRON CAPTURE RADIOISOTOPES; INTERMEDIATE MASS NUCLEI; IODINE ISOTOPES; ISOTOPES; MATERIALS; MEMBRANES; NUCLEI; ODD-EVEN NUCLEI; ORGANIC COMPOUNDS; RADIOISOTOPES; 550201* - Biochemistry- Tracer Techniques

Citation Formats


                    Saboori, A M, Smith, B L, and Agre, P. Polymorphism in the M sub r 32,000 Rh protein purified from Rh(D)-positive and -negative erythrocytes.  United States: N. p., 1988. 
        Web.  doi:10.1073/pnas.85.11.4042.

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                    Saboori, A M, Smith, B L, & Agre, P. Polymorphism in the M sub r 32,000 Rh protein purified from Rh(D)-positive and -negative erythrocytes.  United States.  https://doi.org/10.1073/pnas.85.11.4042

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                    Saboori, A M, Smith, B L, and Agre, P. 1988.  
        "Polymorphism in the M sub r 32,000 Rh protein purified from Rh(D)-positive and -negative erythrocytes".  United States.  https://doi.org/10.1073/pnas.85.11.4042.

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@article{osti_5565401,

  title        = {Polymorphism in the M sub r 32,000 Rh protein purified from Rh(D)-positive and -negative erythrocytes},

  author       = {Saboori, A M and Smith, B L and Agre, P},

  abstractNote = {A M{sub r} 32,000 integral membrane protein has previously been identified on erythrocytes bearing the Rh(D) antigen and is thought to contain the antigenic variations responsible for the different Rh phenotypes. To study it on a biochemical level, a simple large-scale method was developed to purify the M{sub r} 32,000 Rh protein from multiple units of Rh(D)-positive and -negative blood. Erythrocyte membrane vesicles were solubilized in NaDodSO{sub 4}, and a tracer of immunoprecipitated {sup 125}I surface-labeled Rh protein was added. The Rh protein was purified to homogeneity by hydroxylapatite chromatography followed by preparative NaDodSO{sub 4}/PAGE. Approximately 25 nmol of pure Rh protein was recovered from each unit of Rh(D)-positive and -negative blood. Rh protein purified from both Rh phenotypes appeared similar by one-dimensional NaDodSO{sub 4}/PAGE, and the N-terminal amino acid sequences for the first 20 residues were identical. Rh proteins purified from Rh(D)-positive and -negative blood were compared by two-dimensional iodopeptide mapping after {sup 125}I-labeling and {alpha}-chymotrypsin digestion. The peptide maps were very similar. These data indicate that a similar core Rh protein exists in both Rh(D)-positive and -negative erythrocytes, and the Rh proteins from erythrocytes with different Rh phenotypes contain distinct structural polymorphisms.},

  doi          = {10.1073/pnas.85.11.4042},

  url          = {https://www.osti.gov/biblio/5565401},
  journal      = {Proceedings of the National Academy of Sciences of the United States of America; (USA)},

  issn         = {0027-8424},

  number       = ,

  volume       = 85:11,

  place        = {United States},

  year         = {Wed Jun 01 00:00:00 EDT 1988},

  month        = {Wed Jun 01 00:00:00 EDT 1988}

}

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