DOI: 10.1073/pnas.85.14.5026. Purification of ras GTPase activating protein from bovine brain.

Purification of ras GTPase activating protein from bovine brain.

10.1073/pnas.85.14.5026

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

In cytosolic extracts of bovine brain, we detected ras GTPase activating protein (GAP) activity that stimulated the GTP hydrolytic activity of normal c-Ha-ras p21 but not that of the oncogenic [Val12]p21 variant. GAP was purified 19,500-fold by a five-column procedure involving DEAE-Sephacel, Sepharose 6B, orange dye and green dye matrices, and Mono Q resins. A single major protein band of 125 kDa was observed on NaDodSO4/polyacrylamide gels that correlated with the elution of GAP activity on Mono Q. Purified GAP was devoid of inherent GTP hydrolytic activity, suggesting that it was a regulator of ras intrinsic GTPase activity. Under submaximal velocity conditions, the second-order rate constant of GTP hydrolysis at 24 degrees C for p21-GTP + GAP (4.5 X 10(6) M-1.sec-1) was at least 1000-fold greater than that for [Val12]p21-GTP + GAP (less than 3 X 10(3) M-1.sec-1).

Bibliography

Gibbs, J. B., Schaber, M. D., Allard, W. J., Sigal, I. S., & Scolnick, E. M. (1988). Purification of ras GTPase activating protein from bovine brain. Proceedings of the National Academy of Sciences, 85(14), 5026â5030.

Authors 5

J B Gibbs (first)
M D Schaber (additional)
W J Allard (additional)
I S Sigal (additional)
E M Scolnick (additional)

References 0 Referenced 165

None

Dates

Type	When
Created	19 years, 2 months ago (May 31, 2006, 6:38 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 12:56 p.m.)
Indexed	3 weeks, 5 days ago (July 30, 2025, 10:45 a.m.)
Issued	37 years, 1 month ago (July 1, 1988)
Published	37 years, 1 month ago (July 1, 1988)
Published Online	37 years, 1 month ago (July 1, 1988)
Published Print	37 years, 1 month ago (July 1, 1988)

Funders 0

None

BibTeX

@article{Gibbs_1988, title={Purification of ras GTPase activating protein from bovine brain.}, volume={85}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.85.14.5026}, DOI={10.1073/pnas.85.14.5026}, number={14}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Gibbs, J B and Schaber, M D and Allard, W J and Sigal, I S and Scolnick, E M}, year={1988}, month=jul, pages={5026–5030} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        7,
        30
      ]
    ],
    "date-time": "2025-07-30T14:45:06Z",
    "timestamp": 1753886706778
  },
  "reference-count": 0,
  "publisher": "Proceedings of the National Academy of Sciences",
  "issue": "14",
  "content-domain": {
    "domain": [
      "www.pnas.org"
    ],
    "crossmark-restriction": true
  },
  "published-print": {
    "date-parts": [
      [
        1988,
        7
      ]
    ]
  },
  "abstract": "<jats:p>In cytosolic extracts of bovine brain, we detected ras GTPase activating protein (GAP) activity that stimulated the GTP hydrolytic activity of normal c-Ha-ras p21 but not that of the oncogenic [Val12]p21 variant. GAP was purified 19,500-fold by a five-column procedure involving DEAE-Sephacel, Sepharose 6B, orange dye and green dye matrices, and Mono Q resins. A single major protein band of 125 kDa was observed on NaDodSO4/polyacrylamide gels that correlated with the elution of GAP activity on Mono Q. Purified GAP was devoid of inherent GTP hydrolytic activity, suggesting that it was a regulator of ras intrinsic GTPase activity. Under submaximal velocity conditions, the second-order rate constant of GTP hydrolysis at 24 degrees C for p21-GTP + GAP (4.5 X 10(6) M-1.sec-1) was at least 1000-fold greater than that for [Val12]p21-GTP + GAP (less than 3 X 10(3) M-1.sec-1).</jats:p>",
  "DOI": "10.1073/pnas.85.14.5026",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2006,
        5,
        31
      ]
    ],
    "date-time": "2006-05-31T10:38:36Z",
    "timestamp": 1149071916000
  },
  "page": "5026-5030",
  "update-policy": "http://dx.doi.org/10.1073/pnas.cm10313",
  "source": "Crossref",
  "is-referenced-by-count": 165,
  "title": "Purification of ras GTPase activating protein from bovine brain.",
  "prefix": "10.1073",
  "volume": "85",
  "author": [
    {
      "given": "J B",
      "family": "Gibbs",
      "sequence": "first",
      "affiliation": [
        {
          "name": "Department of Molecular Biology, Merck Sharp &amp; Dohme Research Laboratories, West Point, PA 19486."
        }
      ]
    },
    {
      "given": "M D",
      "family": "Schaber",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Molecular Biology, Merck Sharp &amp; Dohme Research Laboratories, West Point, PA 19486."
        }
      ]
    },
    {
      "given": "W J",
      "family": "Allard",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Molecular Biology, Merck Sharp &amp; Dohme Research Laboratories, West Point, PA 19486."
        }
      ]
    },
    {
      "given": "I S",
      "family": "Sigal",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Molecular Biology, Merck Sharp &amp; Dohme Research Laboratories, West Point, PA 19486."
        }
      ]
    },
    {
      "given": "E M",
      "family": "Scolnick",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Molecular Biology, Merck Sharp &amp; Dohme Research Laboratories, West Point, PA 19486."
        }
      ]
    }
  ],
  "member": "341",
  "published-online": {
    "date-parts": [
      [
        1988,
        7
      ]
    ]
  },
  "container-title": "Proceedings of the National Academy of Sciences",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://pnas.org/doi/pdf/10.1073/pnas.85.14.5026",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2022,
        4,
        13
      ]
    ],
    "date-time": "2022-04-13T16:56:51Z",
    "timestamp": 1649869011000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://pnas.org/doi/full/10.1073/pnas.85.14.5026"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        1988,
        7
      ]
    ]
  },
  "references-count": 0,
  "journal-issue": {
    "issue": "14",
    "published-print": {
      "date-parts": [
        [
          1988,
          7
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1073/pnas.85.14.5026"
  ],
  "URL": "http://dx.doi.org/10.1073/pnas.85.14.5026",
  "relation": {},
  "ISSN": [
    "0027-8424",
    "1091-6490"
  ],
  "subject": [],
  "container-title-short": "Proc. Natl. Acad. Sci. U.S.A.",
  "published": {
    "date-parts": [
      [
        1988,
        7
      ]
    ]
  },
  "assertion": [
    {
      "value": "1988-07-01",
      "order": 2,
      "name": "published",
      "label": "Published",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    }
  ]
}