DOI: 10.1111/j.1471-4159.1983.tb04747.x. Affinity‐Purified Anti‐B‐50 Protein Antibody: Interference with the Function of the Phosphoprotein B‐50 in Synaptic Plasma Membranes

Affinity‐Purified Anti‐B‐50 Protein Antibody: Interference with the Function of the Phosphoprotein B‐50 in Synaptic Plasma Membranes

10.1111/j.1471-4159.1983.tb04747.x

Crossref journal-article

Wiley

Journal of Neurochemistry (311)

Abstract

Abstract: Affinity‐purified anti‐B‐50 protein antibodies were used to study the previously proposed relationship of the phosphorylation state of B‐50 protein and polyphosphoinositide metabolism in synaptic plasma membranes. Antibodies were raised against a membrane extract enriched in the B‐50 protein and its adrenocorticotropin‐sensitive protein kinase, obtained from rat brain. Anti‐B‐50 protein immunoglobulins were purified by affinity chromatography on a solid immunosorbent prepared from B‐50 protein isolated by an improved procedure. The purified antibodies reacted only with the B‐50 and B‐60 protein, a proteolysis derivative (of B‐50), as assessed by the sodium dodecyl sulfate‐gel immunoperoxidase method. These antibodies inhibited specifically the endogenous phosphorylation of B‐50 protein in synaptic plasma membranes, without affecting notably the phosphorylation of other membrane proteins. This inhibition was accompanied by changes of the formation of phosphatidylinositol 4,5‐diphosphate and phosphatidic acid in synaptic plasma membranes, whereas formation of phosphatidylinositol 4‐phosphate was not altered. Inhibition by ACTH 1–24 of the endogenous phosphorylation of B‐50 protein in membranes was associated only with an enhancement of the phosphorylation of phosphatidylinositol 4‐phosphate to phosphatidylinositol 4,5‐diphosphate. These data support our hypothesis on the functional interaction of B‐50 protein and phosphatidylinositol 4‐phosphate kinase in rat brain membranes. The evidence shows that purified anti‐B‐50 protein antibodies can be used to probe specifically the function of B‐50 protein in membranes.

Bibliography

Oestreicher, A. B., Van Dongen, C. J., Zwiers, H., & Gispen, W. H. (1983). AffinityâPurified AntiâBâ50 Protein Antibody: Interference with the Function of the Phosphoprotein Bâ50 in Synaptic Plasma Membranes. Journal of Neurochemistry, 41(2), 331â340. Portico.

Authors 4

Anna Beate Oestreicher (first)
Carla J. Van Dongen (additional)
Henk Zwiers (additional)
Willem Hendrik Gispen (additional)

References 39 Referenced 149

{'key': 'e_1_2_2_2_1', 'first-page': '906', 'article-title': 'Calcium‐activated, phospholipid‐dependent protein kinase from rat brain is inhibited by ACTH', 'volume': '7', 'author': 'Aloyo V. J.', 'year': '1981', 'journal-title': 'Soc. Neurosci. Abstr'} / Soc. Neurosci. Abstr / Calcium‐activated, phospholipid‐dependent protein kinase from rat brain is inhibited by ACTH by Aloyo V. J. (1981)
{'key': 'e_1_2_2_3_1', 'first-page': '303', 'volume-title': 'Progress in Brain Research, Vol. 56, Brain Phosphoproteins: Characteristics and Function', 'author': 'Aloyo V. J.', 'year': '1982'} / Progress in Brain Research, Vol. 56, Brain Phosphoproteins: Characteristics and Function by Aloyo V. J. (1982)
10.1016/0006-8993(82)90340-7
10.1007/978-1-4613-4130-7_14
10.1016/0147-7552(77)90037-7
{'key': 'e_1_2_2_7_1', 'first-page': '419', 'volume-title': 'Methods of Enzymology, Vol. 70A', 'author': 'Engvall E.', 'year': '1980'} / Methods of Enzymology, Vol. 70A by Engvall E. (1980)
10.1007/978-1-4684-3503-0_11
10.1126/science.22932
{'key': 'e_1_2_2_10_1', 'first-page': '2208', 'article-title': 'Cyclic nucleotides, phosphorylated proteins and the nervous system', 'volume': '38', 'author': 'Greengard P.', 'year': '1979', 'journal-title': 'Fed. Proc'} / Fed. Proc / Cyclic nucleotides, phosphorylated proteins and the nervous system by Greengard P. (1979)
10.1038/286623a0
10.1042/bj1940283
10.1016/0006-8993(81)90862-3
10.1016/B978-0-12-137020-6.50016-X
10.1016/0005-2736(74)90268-5
10.1111/j.1471-4159.1982.tb03958.x
10.1073/pnas.77.12.7039
10.1016/S0021-9258(19)52451-6
10.1126/science.6162199
10.1016/0968-0004(79)90443-2
10.1016/S0021-9258(19)68675-8 / J. Biol. Chem / Affinity‐purified anti‐protein antibody: Specific inhibitor of phosphorylation of protein I. a synaptic protein by Naito S. (1981)
10.1016/S0361-9230(81)80040-8
{'key': 'e_1_2_2_23_1', 'first-page': '349', 'volume-title': 'Progress in Brain Research, Vol. 55, Chemical Transmission in the Brain', 'author': 'Oestreicher A. B.', 'year': '1982'} / Progress in Brain Research, Vol. 55, Chemical Transmission in the Brain by Oestreicher A. B. (1982)
10.1016/0005-2795(73)90037-8
{'key': 'e_1_2_2_25_1', 'first-page': '1', 'volume-title': 'Progress in Brain Research: Vol. 56: Brain Phosphoproteins: Characteristics and Function', 'author': 'Rodnight R.', 'year': '1982'} / Progress in Brain Research: Vol. 56: Brain Phosphoproteins: Characteristics and Function by Rodnight R. (1982)
10.1111/j.1471-4159.1977.tb07760.x
10.1016/0361-9230(81)90098-8
10.1073/pnas.76.9.4687
{'key': 'e_1_2_2_29_1', 'first-page': '287', 'volume-title': 'Progress in Brain Research: Vol. 56: Brain Phosphoproteins: Characteristics and Function', 'author': 'Takai Y.', 'year': '1982'} / Progress in Brain Research: Vol. 56: Brain Phosphoproteins: Characteristics and Function by Takai Y. (1982)
10.1016/S0021-9258(17)40170-0 / J. Biol. Chem / Adenosine 3′,5′‐monophosphate‐regulated phosphoprotein system of neuronal membranes. 1. Solubilization, purification and some properties of an endogenous phosphoprotein by Ueda T. (1977)
10.1016/0005-2736(76)90219-4
10.1016/0022-1759(77)90116-8
10.1007/BF00966326
10.1007/BF00965607
10.1007/BF00966327
10.1111/j.1471-4159.1979.tb11726.x
10.1111/j.1471-4159.1980.tb11262.x
10.1016/0014-5793(80)80172-4
10.1007/BF00965051
10.1016/S0079-6123(08)63787-5

Dates

Type	When
Created	18 years, 10 months ago (Oct. 5, 2006, 9:42 a.m.)
Deposited	1 year, 10 months ago (Oct. 20, 2023, 7 a.m.)
Indexed	2 months, 3 weeks ago (June 12, 2025, 5:44 a.m.)
Issued	42 years, 7 months ago (Feb. 1, 1983)
Published	42 years, 7 months ago (Feb. 1, 1983)
Published Online	18 years, 10 months ago (Oct. 5, 2006)
Published Print	42 years, 7 months ago (Feb. 1, 1983)

Funders 0

None

BibTeX

@article{Oestreicher_1983, title={Affinity‐Purified Anti‐B‐50 Protein Antibody: Interference with the Function of the Phosphoprotein B‐50 in Synaptic Plasma Membranes}, volume={41}, ISSN={1471-4159}, url={http://dx.doi.org/10.1111/j.1471-4159.1983.tb04747.x}, DOI={10.1111/j.1471-4159.1983.tb04747.x}, number={2}, journal={Journal of Neurochemistry}, publisher={Wiley}, author={Oestreicher, Anna Beate and Van Dongen, Carla J. and Zwiers, Henk and Gispen, Willem Hendrik}, year={1983}, month=feb, pages={331–340} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        6,
        12
      ]
    ],
    "date-time": "2025-06-12T09:44:04Z",
    "timestamp": 1749721444161
  },
  "reference-count": 39,
  "publisher": "Wiley",
  "issue": "2",
  "license": [
    {
      "start": {
        "date-parts": [
          [
            2006,
            10,
            5
          ]
        ],
        "date-time": "2006-10-05T00:00:00Z",
        "timestamp": 1160006400000
      },
      "content-version": "vor",
      "delay-in-days": 8647,
      "URL": "http://onlinelibrary.wiley.com/termsAndConditions#vor"
    }
  ],
  "content-domain": {
    "domain": [],
    "crossmark-restriction": false
  },
  "published-print": {
    "date-parts": [
      [
        1983,
        2
      ]
    ]
  },
  "abstract": "<jats:p><jats:bold>Abstract: </jats:bold> Affinity\u2010purified anti\u2010B\u201050 protein antibodies were used to study the previously proposed relationship of the phosphorylation state of B\u201050 protein and polyphosphoinositide metabolism in synaptic plasma membranes. Antibodies were raised against a membrane extract enriched in the B\u201050 protein and its adrenocorticotropin\u2010sensitive protein kinase, obtained from rat brain. Anti\u2010B\u201050 protein immunoglobulins were purified by affinity chromatography on a solid immunosorbent prepared from B\u201050 protein isolated by an improved procedure. The purified antibodies reacted only with the B\u201050 and B\u201060 protein, a proteolysis derivative (of B\u201050), as assessed by the sodium dodecyl sulfate\u2010gel immunoperoxidase method. These antibodies inhibited specifically the endogenous phosphorylation of B\u201050 protein in synaptic plasma membranes, without affecting notably the phosphorylation of other membrane proteins. This inhibition was accompanied by changes of the formation of phosphatidylinositol 4,5\u2010diphosphate and phosphatidic acid in synaptic plasma membranes, whereas formation of phosphatidylinositol 4\u2010phosphate was not altered. Inhibition by ACTH <jats:sub>1\u201324</jats:sub> of the endogenous phosphorylation of B\u201050 protein in membranes was associated only with an enhancement of the phosphorylation of phosphatidylinositol 4\u2010phosphate to phosphatidylinositol 4,5\u2010diphosphate. These data support our hypothesis on the functional interaction of B\u201050 protein and phosphatidylinositol 4\u2010phosphate kinase in rat brain membranes. The evidence shows that purified anti\u2010B\u201050 protein antibodies can be used to probe specifically the function of B\u201050 protein in membranes.</jats:p>",
  "DOI": "10.1111/j.1471-4159.1983.tb04747.x",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2006,
        10,
        5
      ]
    ],
    "date-time": "2006-10-05T13:42:18Z",
    "timestamp": 1160055738000
  },
  "page": "331-340",
  "source": "Crossref",
  "is-referenced-by-count": 149,
  "title": "Affinity\u2010Purified Anti\u2010B\u201050 Protein Antibody: Interference with the Function of the Phosphoprotein B\u201050 in Synaptic Plasma Membranes",
  "prefix": "10.1111",
  "volume": "41",
  "author": [
    {
      "given": "Anna Beate",
      "family": "Oestreicher",
      "sequence": "first",
      "affiliation": []
    },
    {
      "given": "Carla J.",
      "family": "Van Dongen",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Henk",
      "family": "Zwiers",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Willem Hendrik",
      "family": "Gispen",
      "sequence": "additional",
      "affiliation": []
    }
  ],
  "member": "311",
  "published-online": {
    "date-parts": [
      [
        2006,
        10,
        5
      ]
    ]
  },
  "reference": [
    {
      "key": "e_1_2_2_2_1",
      "first-page": "906",
      "article-title": "Calcium\u2010activated, phospholipid\u2010dependent protein kinase from rat brain is inhibited by ACTH",
      "volume": "7",
      "author": "Aloyo V. J.",
      "year": "1981",
      "journal-title": "Soc. Neurosci. Abstr"
    },
    {
      "key": "e_1_2_2_3_1",
      "first-page": "303",
      "volume-title": "Progress in Brain Research, Vol. 56, Brain Phosphoproteins: Characteristics and Function",
      "author": "Aloyo V. J.",
      "year": "1982"
    },
    {
      "key": "e_1_2_2_4_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0006-8993(82)90340-7"
    },
    {
      "key": "e_1_2_2_5_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/978-1-4613-4130-7_14"
    },
    {
      "key": "e_1_2_2_6_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0147-7552(77)90037-7"
    },
    {
      "key": "e_1_2_2_7_1",
      "first-page": "419",
      "volume-title": "Methods of Enzymology, Vol. 70A",
      "author": "Engvall E.",
      "year": "1980"
    },
    {
      "key": "e_1_2_2_8_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/978-1-4684-3503-0_11"
    },
    {
      "key": "e_1_2_2_9_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.22932"
    },
    {
      "key": "e_1_2_2_10_1",
      "first-page": "2208",
      "article-title": "Cyclic nucleotides, phosphorylated proteins and the nervous system",
      "volume": "38",
      "author": "Greengard P.",
      "year": "1979",
      "journal-title": "Fed. Proc"
    },
    {
      "key": "e_1_2_2_11_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/286623a0"
    },
    {
      "key": "e_1_2_2_12_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1042/bj1940283"
    },
    {
      "key": "e_1_2_2_13_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0006-8993(81)90862-3"
    },
    {
      "key": "e_1_2_2_14_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/B978-0-12-137020-6.50016-X"
    },
    {
      "key": "e_1_2_2_15_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0005-2736(74)90268-5"
    },
    {
      "key": "e_1_2_2_16_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1471-4159.1982.tb03958.x"
    },
    {
      "key": "e_1_2_2_17_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.77.12.7039"
    },
    {
      "key": "e_1_2_2_18_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0021-9258(19)52451-6"
    },
    {
      "key": "e_1_2_2_19_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.6162199"
    },
    {
      "key": "e_1_2_2_20_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0968-0004(79)90443-2"
    },
    {
      "key": "e_1_2_2_21_1",
      "doi-asserted-by": "crossref",
      "first-page": "10657",
      "DOI": "10.1016/S0021-9258(19)68675-8",
      "article-title": "Affinity\u2010purified anti\u2010protein antibody: Specific inhibitor of phosphorylation of protein I. a synaptic protein",
      "volume": "256",
      "author": "Naito S.",
      "year": "1981",
      "journal-title": "J. Biol. Chem"
    },
    {
      "key": "e_1_2_2_22_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0361-9230(81)80040-8"
    },
    {
      "key": "e_1_2_2_23_1",
      "first-page": "349",
      "volume-title": "Progress in Brain Research, Vol. 55, Chemical Transmission in the Brain",
      "author": "Oestreicher A. B.",
      "year": "1982"
    },
    {
      "key": "e_1_2_2_24_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0005-2795(73)90037-8"
    },
    {
      "key": "e_1_2_2_25_1",
      "first-page": "1",
      "volume-title": "Progress in Brain Research: Vol. 56: Brain Phosphoproteins: Characteristics and Function",
      "author": "Rodnight R.",
      "year": "1982"
    },
    {
      "key": "e_1_2_2_26_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1471-4159.1977.tb07760.x"
    },
    {
      "key": "e_1_2_2_27_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0361-9230(81)90098-8"
    },
    {
      "key": "e_1_2_2_28_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.76.9.4687"
    },
    {
      "key": "e_1_2_2_29_1",
      "first-page": "287",
      "volume-title": "Progress in Brain Research: Vol. 56: Brain Phosphoproteins: Characteristics and Function",
      "author": "Takai Y.",
      "year": "1982"
    },
    {
      "key": "e_1_2_2_30_1",
      "doi-asserted-by": "crossref",
      "first-page": "5155",
      "DOI": "10.1016/S0021-9258(17)40170-0",
      "article-title": "Adenosine 3\u2032,5\u2032\u2010monophosphate\u2010regulated phosphoprotein system of neuronal membranes. 1. Solubilization, purification and some properties of an endogenous phosphoprotein",
      "volume": "252",
      "author": "Ueda T.",
      "year": "1977",
      "journal-title": "J. Biol. Chem"
    },
    {
      "key": "e_1_2_2_31_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0005-2736(76)90219-4"
    },
    {
      "key": "e_1_2_2_32_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0022-1759(77)90116-8"
    },
    {
      "key": "e_1_2_2_33_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/BF00966326"
    },
    {
      "key": "e_1_2_2_34_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/BF00965607"
    },
    {
      "key": "e_1_2_2_35_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/BF00966327"
    },
    {
      "key": "e_1_2_2_36_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1471-4159.1979.tb11726.x"
    },
    {
      "key": "e_1_2_2_37_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1111/j.1471-4159.1980.tb11262.x"
    },
    {
      "key": "e_1_2_2_38_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0014-5793(80)80172-4"
    },
    {
      "key": "e_1_2_2_39_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/BF00965051"
    },
    {
      "key": "e_1_2_2_40_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0079-6123(08)63787-5"
    }
  ],
  "container-title": "Journal of Neurochemistry",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1111%2Fj.1471-4159.1983.tb04747.x",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "text-mining"
    },
    {
      "URL": "https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1471-4159.1983.tb04747.x",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2023,
        10,
        20
      ]
    ],
    "date-time": "2023-10-20T11:00:19Z",
    "timestamp": 1697799619000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://onlinelibrary.wiley.com/doi/10.1111/j.1471-4159.1983.tb04747.x"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        1983,
        2
      ]
    ]
  },
  "references-count": 39,
  "journal-issue": {
    "issue": "2",
    "published-print": {
      "date-parts": [
        [
          1983,
          2
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1111/j.1471-4159.1983.tb04747.x"
  ],
  "URL": "http://dx.doi.org/10.1111/j.1471-4159.1983.tb04747.x",
  "relation": {},
  "ISSN": [
    "0022-3042",
    "1471-4159"
  ],
  "subject": [],
  "container-title-short": "Journal of Neurochemistry",
  "published": {
    "date-parts": [
      [
        1983,
        2
      ]
    ]
  }
}