DOI: 10.1110/ps.9.4.647. Mutational analysis of the major coat protein of M13 identifies residues that control protein display

Mutational analysis of the major coat protein of M13 identifies residues that control protein display

10.1110/ps.9.4.647

Crossref journal-article

Wiley

Protein Science (311)

Abstract

AbstractWe have reported variants of the M13 bacteriophage major coat protein (P8) that enable high copy display of monomeric and oligomeric proteins, such as human growth hormone and steptavidin, on the surface of phage particles (Sidhu SS, Weiss GA, Wells JA. 2000. High copy display of large proteins on phage for functional selections. J Mol Biol 296:487–495). Here, we explore how an optimized P8 variant (opti‐P8) could evolve the ability to efficiently display a protein fused to its N‐terminus. Reversion of individual opti‐P8 residues back to the wild‐type P8 residue identifies a limited set of hydrophobic residues responsible for the high copy protein display. These hydrophobic amino acids bracket a conserved hydrophobic face on the P8 alpha helix thought to be in contact with the phage coat. Mutations additively combine to promote high copy protein display, which was further enhanced by optimization of the linker between the phage coat and the fusion protein. These data are consistent with a model in which protein displayenhancing mutations allow for better packing of the fusion protein into the phage coat. The high tolerance for phage coat protein mutations observed here suggests that filamentous phage coat proteins could readily evolve new capabilities.

Authors 3

Gregory A. Weiss (first)
James A. Wells (additional)
Sachdev S. Sidhu (additional)

References 26 Referenced 25

{'key': 'e_1_2_1_2_1', 'first-page': '376', 'article-title': 'XL1‐Blue: A high efficiency plasmid transfonning recA Escherichia coli strain with beta‐galactosidase selection', 'volume': '5', 'author': 'Bullock WO', 'year': '1987', 'journal-title': 'Biotechniques'} / Biotechniques / XL1‐Blue: A high efficiency plasmid transfonning recA Escherichia coli strain with beta‐galactosidase selection by Bullock WO (1987)
10.1016/S0022-1759(98)00094-5
10.1016/0167-7799(94)90079-5
10.1016/S0021-9258(19)39741-8
10.1006/jmbi.1998.1778
10.1006/jmbi.1995.0264
10.1016/0022-2836(92)90636-X
10.1073/pnas.88.10.4363
10.1038/20715
10.1016/0378-1119(94)00897-2
10.1016/0076-6879(87)54085-X
{'key': 'e_1_2_1_13_1', 'first-page': '249', 'article-title': 'Phage display of peptide libraries on protein scaffolds', 'volume': '87', 'author': 'Lowman HB', 'year': '1998', 'journal-title': 'Methods Mol Biol'} / Methods Mol Biol / Phage display of peptide libraries on protein scaffolds by Lowman HB (1998)
10.1006/jmbi.1996.0378
10.1016/S0022-2836(05)80032-4
10.1016/S1380-2933(96)00049-8
10.1002/prot.340110407
10.1006/jmbi.1998.1860
10.1074/jbc.272.33.20595
10.1006/jmbi.1993.1320
SidhuSS LowmanHB CunninghamBC WellsJA.2000a. Phage display of novel binding peptides.Methods Enzymol.Forthcoming.
10.1006/jmbi.1999.3465
10.1126/science.4001944
10.1021/cr960065d
10.1006/jmbi.1998.2081
10.1006/jmbi.1995.0469
10.1139/w98-015

Dates

Type	When
Created	14 years, 10 months ago (Oct. 1, 2010, 9:28 a.m.)
Deposited	1 year, 10 months ago (Oct. 29, 2023, 5:32 a.m.)
Indexed	1 year ago (Aug. 7, 2024, 9:53 a.m.)
Issued	25 years, 7 months ago (Jan. 1, 2000)
Published	25 years, 7 months ago (Jan. 1, 2000)
Published Online	16 years, 8 months ago (Dec. 31, 2008)
Published Print	25 years, 7 months ago (Jan. 1, 2000)

Funders 0

None

BibTeX

@article{Weiss_2000, title={Mutational analysis of the major coat protein of M13 identifies residues that control protein display}, volume={9}, ISSN={1469-896X}, url={http://dx.doi.org/10.1110/ps.9.4.647}, DOI={10.1110/ps.9.4.647}, number={4}, journal={Protein Science}, publisher={Wiley}, author={Weiss, Gregory A. and Wells, James A. and Sidhu, Sachdev S.}, year={2000}, month=jan, pages={647–654} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2024,
        8,
        7
      ]
    ],
    "date-time": "2024-08-07T13:53:34Z",
    "timestamp": 1723038814638
  },
  "reference-count": 26,
  "publisher": "Wiley",
  "issue": "4",
  "license": [
    {
      "start": {
        "date-parts": [
          [
            2008,
            12,
            31
          ]
        ],
        "date-time": "2008-12-31T00:00:00Z",
        "timestamp": 1230681600000
      },
      "content-version": "vor",
      "delay-in-days": 3287,
      "URL": "http://onlinelibrary.wiley.com/termsAndConditions#vor"
    }
  ],
  "content-domain": {
    "domain": [],
    "crossmark-restriction": false
  },
  "published-print": {
    "date-parts": [
      [
        2000,
        1
      ]
    ]
  },
  "abstract": "<jats:title>Abstract</jats:title><jats:p>We have reported variants of the M13 bacteriophage major coat protein (P8) that enable high copy display of monomeric and oligomeric proteins, such as human growth hormone and steptavidin, on the surface of phage particles (Sidhu SS, Weiss GA, Wells JA. 2000. High copy display of large proteins on phage for functional selections. <jats:italic>J Mol Biol 296</jats:italic>:487\u2013495). Here, we explore how an optimized P8 variant (opti\u2010P8) could evolve the ability to efficiently display a protein fused to its N\u2010terminus. Reversion of individual opti\u2010P8 residues back to the wild\u2010type P8 residue identifies a limited set of hydrophobic residues responsible for the high copy protein display. These hydrophobic amino acids bracket a conserved hydrophobic face on the P8 alpha helix thought to be in contact with the phage coat. Mutations additively combine to promote high copy protein display, which was further enhanced by optimization of the linker between the phage coat and the fusion protein. These data are consistent with a model in which protein displayenhancing mutations allow for better packing of the fusion protein into the phage coat. The high tolerance for phage coat protein mutations observed here suggests that filamentous phage coat proteins could readily evolve new capabilities.</jats:p>",
  "DOI": "10.1110/ps.9.4.647",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2010,
        10,
        1
      ]
    ],
    "date-time": "2010-10-01T13:28:04Z",
    "timestamp": 1285939684000
  },
  "page": "647-654",
  "source": "Crossref",
  "is-referenced-by-count": 25,
  "title": "Mutational analysis of the major coat protein of M13 identifies residues that control protein display",
  "prefix": "10.1002",
  "volume": "9",
  "author": [
    {
      "given": "Gregory A.",
      "family": "Weiss",
      "sequence": "first",
      "affiliation": []
    },
    {
      "given": "James A.",
      "family": "Wells",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Sachdev S.",
      "family": "Sidhu",
      "sequence": "additional",
      "affiliation": []
    }
  ],
  "member": "311",
  "published-online": {
    "date-parts": [
      [
        2008,
        12,
        31
      ]
    ]
  },
  "reference": [
    {
      "key": "e_1_2_1_2_1",
      "first-page": "376",
      "article-title": "XL1\u2010Blue: A high efficiency plasmid transfonning recA Escherichia coli strain with beta\u2010galactosidase selection",
      "volume": "5",
      "author": "Bullock WO",
      "year": "1987",
      "journal-title": "Biotechniques"
    },
    {
      "key": "e_1_2_1_3_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0022-1759(98)00094-5"
    },
    {
      "key": "e_1_2_1_4_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0167-7799(94)90079-5"
    },
    {
      "key": "e_1_2_1_5_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0021-9258(19)39741-8"
    },
    {
      "key": "e_1_2_1_6_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/jmbi.1998.1778"
    },
    {
      "key": "e_1_2_1_7_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/jmbi.1995.0264"
    },
    {
      "key": "e_1_2_1_8_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0022-2836(92)90636-X"
    },
    {
      "key": "e_1_2_1_9_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.88.10.4363"
    },
    {
      "key": "e_1_2_1_10_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/20715"
    },
    {
      "key": "e_1_2_1_11_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0378-1119(94)00897-2"
    },
    {
      "key": "e_1_2_1_12_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0076-6879(87)54085-X"
    },
    {
      "key": "e_1_2_1_13_1",
      "first-page": "249",
      "article-title": "Phage display of peptide libraries on protein scaffolds",
      "volume": "87",
      "author": "Lowman HB",
      "year": "1998",
      "journal-title": "Methods Mol Biol"
    },
    {
      "key": "e_1_2_1_14_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/jmbi.1996.0378"
    },
    {
      "key": "e_1_2_1_15_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0022-2836(05)80032-4"
    },
    {
      "key": "e_1_2_1_16_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S1380-2933(96)00049-8"
    },
    {
      "key": "e_1_2_1_17_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/prot.340110407"
    },
    {
      "key": "e_1_2_1_18_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/jmbi.1998.1860"
    },
    {
      "key": "e_1_2_1_19_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.272.33.20595"
    },
    {
      "key": "e_1_2_1_20_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/jmbi.1993.1320"
    },
    {
      "key": "e_1_2_1_21_1",
      "unstructured": "SidhuSS LowmanHB CunninghamBC WellsJA.2000a. Phage display of novel binding peptides.Methods Enzymol.Forthcoming."
    },
    {
      "key": "e_1_2_1_22_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/jmbi.1999.3465"
    },
    {
      "key": "e_1_2_1_23_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.4001944"
    },
    {
      "key": "e_1_2_1_24_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1021/cr960065d"
    },
    {
      "key": "e_1_2_1_25_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/jmbi.1998.2081"
    },
    {
      "key": "e_1_2_1_26_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/jmbi.1995.0469"
    },
    {
      "key": "e_1_2_1_27_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1139/w98-015"
    }
  ],
  "container-title": "Protein Science",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1110%2Fps.9.4.647",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "text-mining"
    },
    {
      "URL": "https://onlinelibrary.wiley.com/doi/pdf/10.1110/ps.9.4.647",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2023,
        10,
        29
      ]
    ],
    "date-time": "2023-10-29T09:32:12Z",
    "timestamp": 1698571932000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://onlinelibrary.wiley.com/doi/10.1110/ps.9.4.647"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        2000,
        1
      ]
    ]
  },
  "references-count": 26,
  "journal-issue": {
    "issue": "4",
    "published-print": {
      "date-parts": [
        [
          2000,
          1
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1110/ps.9.4.647"
  ],
  "URL": "http://dx.doi.org/10.1110/ps.9.4.647",
  "relation": {},
  "ISSN": [
    "0961-8368",
    "1469-896X"
  ],
  "subject": [],
  "container-title-short": "Protein Science",
  "published": {
    "date-parts": [
      [
        2000,
        1
      ]
    ]
  }
}