Eintrag weiter verarbeiten
Trapping a 96° domain rotation in two distinct conformations by engineered disulfide bridges
Gespeichert in:
| Zeitschriftentitel: | Protein Science |
|---|---|
| Personen und Körperschaften: | , , |
| In: | Protein Science, 13, 2004, 7, S. 1811-1822 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
Wiley
|
| Schlagwörter: |
| author_facet |
Schultz‐Heienbrok, Robert Maier, Timm Sträter, Norbert Schultz‐Heienbrok, Robert Maier, Timm Sträter, Norbert |
|---|---|
| author |
Schultz‐Heienbrok, Robert Maier, Timm Sträter, Norbert |
| spellingShingle |
Schultz‐Heienbrok, Robert Maier, Timm Sträter, Norbert Protein Science Trapping a 96° domain rotation in two distinct conformations by engineered disulfide bridges Molecular Biology Biochemistry |
| author_sort |
schultz‐heienbrok, robert |
| spelling |
Schultz‐Heienbrok, Robert Maier, Timm Sträter, Norbert 0961-8368 1469-896X Wiley Molecular Biology Biochemistry http://dx.doi.org/10.1110/ps.04629604 <jats:title>Abstract</jats:title><jats:p>Engineering disulfide bridges is a common technique to lock a protein movement in a defined conformational state. We have designed two double mutants of <jats:italic>Escherichia coli</jats:italic> 5′‐nucleotidase to trap the enzyme in both an open (S228C, P513C) and a closed (P90C, L424C) conformation by the formation of disulfide bridges. The mutant proteins have been expressed, purified, and crystallized, to structurally characterize the designed variants. The S228C, P513C is a double mutant crystallized in two different crystal forms with three independent conformers, which differ from each other by a rotation of up to 12° of the C‐terminal domain with respect to the N‐terminal domain. This finding, as well as an analysis of the domain motion in the crystal, indicates that the enzyme still exhibits considerable residual domain flexibility. In the double mutant that was designed to trap the enzyme in the closed conformation, the structure analysis reveals an unexpected intermediate conformation along the 96° rotation trajectory between the open and closed enzyme forms. A comparison of the five independent conformers analyzed in this study shows that the domain movement of the variant enzymes is characterized by a sliding movement of the residues of the domain interface along the interface, which is in contrast to a classical closure motion where the residues of the domain interface move perpendicular to the interface.</jats:p> Trapping a 96° domain rotation in two distinct conformations by engineered disulfide bridges Protein Science |
| doi_str_mv |
10.1110/ps.04629604 |
| facet_avail |
Online Free |
| finc_class_facet |
Chemie und Pharmazie Biologie |
| format |
ElectronicArticle |
| fullrecord |
blob:ai-49-aHR0cDovL2R4LmRvaS5vcmcvMTAuMTExMC9wcy4wNDYyOTYwNA |
| id |
ai-49-aHR0cDovL2R4LmRvaS5vcmcvMTAuMTExMC9wcy4wNDYyOTYwNA |
| institution |
DE-Zi4 DE-Gla1 DE-15 DE-Pl11 DE-Rs1 DE-14 DE-105 DE-Ch1 DE-L229 DE-D275 DE-Bn3 DE-Brt1 DE-Zwi2 DE-D161 |
| imprint |
Wiley, 2004 |
| imprint_str_mv |
Wiley, 2004 |
| issn |
1469-896X 0961-8368 |
| issn_str_mv |
1469-896X 0961-8368 |
| language |
English |
| mega_collection |
Wiley (CrossRef) |
| match_str |
schultzheienbrok2004trappinga96degdomainrotationintwodistinctconformationsbyengineereddisulfidebridges |
| publishDateSort |
2004 |
| publisher |
Wiley |
| recordtype |
ai |
| record_format |
ai |
| series |
Protein Science |
| source_id |
49 |
| title |
Trapping a 96° domain rotation in two distinct conformations by engineered disulfide bridges |
| title_unstemmed |
Trapping a 96° domain rotation in two distinct conformations by engineered disulfide bridges |
| title_full |
Trapping a 96° domain rotation in two distinct conformations by engineered disulfide bridges |
| title_fullStr |
Trapping a 96° domain rotation in two distinct conformations by engineered disulfide bridges |
| title_full_unstemmed |
Trapping a 96° domain rotation in two distinct conformations by engineered disulfide bridges |
| title_short |
Trapping a 96° domain rotation in two distinct conformations by engineered disulfide bridges |
| title_sort |
trapping a 96° domain rotation in two distinct conformations by engineered disulfide bridges |
| topic |
Molecular Biology Biochemistry |
| url |
http://dx.doi.org/10.1110/ps.04629604 |
| publishDate |
2004 |
| physical |
1811-1822 |
| description |
<jats:title>Abstract</jats:title><jats:p>Engineering disulfide bridges is a common technique to lock a protein movement in a defined conformational state. We have designed two double mutants of <jats:italic>Escherichia coli</jats:italic> 5′‐nucleotidase to trap the enzyme in both an open (S228C, P513C) and a closed (P90C, L424C) conformation by the formation of disulfide bridges. The mutant proteins have been expressed, purified, and crystallized, to structurally characterize the designed variants. The S228C, P513C is a double mutant crystallized in two different crystal forms with three independent conformers, which differ from each other by a rotation of up to 12° of the C‐terminal domain with respect to the N‐terminal domain. This finding, as well as an analysis of the domain motion in the crystal, indicates that the enzyme still exhibits considerable residual domain flexibility. In the double mutant that was designed to trap the enzyme in the closed conformation, the structure analysis reveals an unexpected intermediate conformation along the 96° rotation trajectory between the open and closed enzyme forms. A comparison of the five independent conformers analyzed in this study shows that the domain movement of the variant enzymes is characterized by a sliding movement of the residues of the domain interface along the interface, which is in contrast to a classical closure motion where the residues of the domain interface move perpendicular to the interface.</jats:p> |
| container_issue |
7 |
| container_start_page |
1811 |
| container_title |
Protein Science |
| container_volume |
13 |
| format_de105 |
Article, E-Article |
| format_de14 |
Article, E-Article |
| format_de15 |
Article, E-Article |
| format_de520 |
Article, E-Article |
| format_de540 |
Article, E-Article |
| format_dech1 |
Article, E-Article |
| format_ded117 |
Article, E-Article |
| format_degla1 |
E-Article |
| format_del152 |
Buch |
| format_del189 |
Article, E-Article |
| format_dezi4 |
Article |
| format_dezwi2 |
Article, E-Article |
| format_finc |
Article, E-Article |
| format_nrw |
Article, E-Article |
| _version_ |
1792336941577928709 |
| geogr_code |
not assigned |
| last_indexed |
2024-03-01T15:08:09.65Z |
| geogr_code_person |
not assigned |
| openURL |
url_ver=Z39.88-2004&ctx_ver=Z39.88-2004&ctx_enc=info%3Aofi%2Fenc%3AUTF-8&rfr_id=info%3Asid%2Fvufind.svn.sourceforge.net%3Agenerator&rft.title=Trapping+a+96%C2%B0+domain+rotation+in+two+distinct+conformations+by+engineered+disulfide+bridges&rft.date=2004-07-01&genre=article&issn=1469-896X&volume=13&issue=7&spage=1811&epage=1822&pages=1811-1822&jtitle=Protein+Science&atitle=Trapping+a+96%C2%B0+domain+rotation+in+two+distinct+conformations+by+engineered+disulfide+bridges&aulast=Str%C3%A4ter&aufirst=Norbert&rft_id=info%3Adoi%2F10.1110%2Fps.04629604&rft.language%5B0%5D=eng |
| SOLR | |
| _version_ | 1792336941577928709 |
| author | Schultz‐Heienbrok, Robert, Maier, Timm, Sträter, Norbert |
| author_facet | Schultz‐Heienbrok, Robert, Maier, Timm, Sträter, Norbert, Schultz‐Heienbrok, Robert, Maier, Timm, Sträter, Norbert |
| author_sort | schultz‐heienbrok, robert |
| container_issue | 7 |
| container_start_page | 1811 |
| container_title | Protein Science |
| container_volume | 13 |
| description | <jats:title>Abstract</jats:title><jats:p>Engineering disulfide bridges is a common technique to lock a protein movement in a defined conformational state. We have designed two double mutants of <jats:italic>Escherichia coli</jats:italic> 5′‐nucleotidase to trap the enzyme in both an open (S228C, P513C) and a closed (P90C, L424C) conformation by the formation of disulfide bridges. The mutant proteins have been expressed, purified, and crystallized, to structurally characterize the designed variants. The S228C, P513C is a double mutant crystallized in two different crystal forms with three independent conformers, which differ from each other by a rotation of up to 12° of the C‐terminal domain with respect to the N‐terminal domain. This finding, as well as an analysis of the domain motion in the crystal, indicates that the enzyme still exhibits considerable residual domain flexibility. In the double mutant that was designed to trap the enzyme in the closed conformation, the structure analysis reveals an unexpected intermediate conformation along the 96° rotation trajectory between the open and closed enzyme forms. A comparison of the five independent conformers analyzed in this study shows that the domain movement of the variant enzymes is characterized by a sliding movement of the residues of the domain interface along the interface, which is in contrast to a classical closure motion where the residues of the domain interface move perpendicular to the interface.</jats:p> |
| doi_str_mv | 10.1110/ps.04629604 |
| facet_avail | Online, Free |
| finc_class_facet | Chemie und Pharmazie, Biologie |
| format | ElectronicArticle |
| format_de105 | Article, E-Article |
| format_de14 | Article, E-Article |
| format_de15 | Article, E-Article |
| format_de520 | Article, E-Article |
| format_de540 | Article, E-Article |
| format_dech1 | Article, E-Article |
| format_ded117 | Article, E-Article |
| format_degla1 | E-Article |
| format_del152 | Buch |
| format_del189 | Article, E-Article |
| format_dezi4 | Article |
| format_dezwi2 | Article, E-Article |
| format_finc | Article, E-Article |
| format_nrw | Article, E-Article |
| geogr_code | not assigned |
| geogr_code_person | not assigned |
| id | ai-49-aHR0cDovL2R4LmRvaS5vcmcvMTAuMTExMC9wcy4wNDYyOTYwNA |
| imprint | Wiley, 2004 |
| imprint_str_mv | Wiley, 2004 |
| institution | DE-Zi4, DE-Gla1, DE-15, DE-Pl11, DE-Rs1, DE-14, DE-105, DE-Ch1, DE-L229, DE-D275, DE-Bn3, DE-Brt1, DE-Zwi2, DE-D161 |
| issn | 1469-896X, 0961-8368 |
| issn_str_mv | 1469-896X, 0961-8368 |
| language | English |
| last_indexed | 2024-03-01T15:08:09.65Z |
| match_str | schultzheienbrok2004trappinga96degdomainrotationintwodistinctconformationsbyengineereddisulfidebridges |
| mega_collection | Wiley (CrossRef) |
| physical | 1811-1822 |
| publishDate | 2004 |
| publishDateSort | 2004 |
| publisher | Wiley |
| record_format | ai |
| recordtype | ai |
| series | Protein Science |
| source_id | 49 |
| spelling | Schultz‐Heienbrok, Robert Maier, Timm Sträter, Norbert 0961-8368 1469-896X Wiley Molecular Biology Biochemistry http://dx.doi.org/10.1110/ps.04629604 <jats:title>Abstract</jats:title><jats:p>Engineering disulfide bridges is a common technique to lock a protein movement in a defined conformational state. We have designed two double mutants of <jats:italic>Escherichia coli</jats:italic> 5′‐nucleotidase to trap the enzyme in both an open (S228C, P513C) and a closed (P90C, L424C) conformation by the formation of disulfide bridges. The mutant proteins have been expressed, purified, and crystallized, to structurally characterize the designed variants. The S228C, P513C is a double mutant crystallized in two different crystal forms with three independent conformers, which differ from each other by a rotation of up to 12° of the C‐terminal domain with respect to the N‐terminal domain. This finding, as well as an analysis of the domain motion in the crystal, indicates that the enzyme still exhibits considerable residual domain flexibility. In the double mutant that was designed to trap the enzyme in the closed conformation, the structure analysis reveals an unexpected intermediate conformation along the 96° rotation trajectory between the open and closed enzyme forms. A comparison of the five independent conformers analyzed in this study shows that the domain movement of the variant enzymes is characterized by a sliding movement of the residues of the domain interface along the interface, which is in contrast to a classical closure motion where the residues of the domain interface move perpendicular to the interface.</jats:p> Trapping a 96° domain rotation in two distinct conformations by engineered disulfide bridges Protein Science |
| spellingShingle | Schultz‐Heienbrok, Robert, Maier, Timm, Sträter, Norbert, Protein Science, Trapping a 96° domain rotation in two distinct conformations by engineered disulfide bridges, Molecular Biology, Biochemistry |
| title | Trapping a 96° domain rotation in two distinct conformations by engineered disulfide bridges |
| title_full | Trapping a 96° domain rotation in two distinct conformations by engineered disulfide bridges |
| title_fullStr | Trapping a 96° domain rotation in two distinct conformations by engineered disulfide bridges |
| title_full_unstemmed | Trapping a 96° domain rotation in two distinct conformations by engineered disulfide bridges |
| title_short | Trapping a 96° domain rotation in two distinct conformations by engineered disulfide bridges |
| title_sort | trapping a 96° domain rotation in two distinct conformations by engineered disulfide bridges |
| title_unstemmed | Trapping a 96° domain rotation in two distinct conformations by engineered disulfide bridges |
| topic | Molecular Biology, Biochemistry |
| url | http://dx.doi.org/10.1110/ps.04629604 |