Crystallization and preliminary X-ray diffraction analysis of human adenovirus

V. S. Reddy; S. K. Natchiar; L. Gritton; T. M. Mullen; P. L. Stewart; G. R. Nemerow

doi:10.1016/j.virol.2010.03.028

Crystallization and preliminary X-ray diffraction analysis of human adenovirus

V. S. Reddy, S. K. Natchiar, L. Gritton, T. M. Mullen, P. L. Stewart, G. R. Nemerow

Research output: Contribution to journal › Article › peer-review

20 Scopus citations

Abstract

Replication-defective and conditionally replicating adenovirus (AdV) vectors are currently being utilized in ~25% of human gene transfer clinical trials. Unfortunately, progress in vector development has been hindered by a lack of accurate structural information. Here we describe the crystallization and preliminary X-ray diffraction analysis of a HAdV5 vector that displays a short flexible fiber derived from HAdV35. Crystals of Ad35F were grown in 100mM HEPES pH 7.0, 200mM Ca(OAc)₂, 14% PEG 550 MME, 15% glycerol in 100mM Tris-HCl 8.5. Freshly grown crystals diffracted well to 4.5Å resolution and weakly to 3.5Å at synchrotron sources. HAdV crystals belong to space group P1 with unit cell parameters a=854.03Å, b=855.17Å, c=865.24Å, α=119.57°, Β=91.71°, γ=118.08° with a single particle in the unit cell. Self-rotation and locked-rotation function analysis allowed the determination of the particle orientation. Molecular replacement, density modification and phase-extension procedures are being employed for structure determination.

Original language	English (US)
Pages (from-to)	209-214
Number of pages	6
Journal	Virology
Volume	402
Issue number	1
DOIs	https://doi.org/10.1016/j.virol.2010.03.028
State	Published - Jun 2010
Externally published	Yes

Bibliographical note

Funding Information:
The authors thank Professor Jack Johnson of TSRI for helpful discussions on aspects of crystallization and data collection. The authors also acknowledge Dr. Sangita Venkataraman for her help with the crystallization setups of HAdV, Dr. Irimpan Mathews of SSRL for his initial help with the data collection and Drs. Wladek Minor and Iwona Minor for generously providing the larger version of SCALEPACK program. We also gratefully acknowledge Drs. Robert Fischetti, Nukri Sanishvili and other members at GM/CA CAT for helpful discussions and technical support. GM/CA CAT has been funded in whole or in part with Federal funds from the National Cancer Institute ( Y1-CO-1020 ) and the National Institute of General Medical Science ( Y1-GM-1104 ). The use of the Advance Photon Source was supported by the U.S. Department of Energy, Basic Energy Sciences, Office of Science (contract No. DE-AC02-06CH11357).

Funding Information:
This work was supported by NIH grants R01 AI070771 to V.S.R and EY011431 and HL54352 to GRN. This is manuscript #20482 at The Scripps Research Institute.

Keywords

Capsid structure
Crystallization
Human adenovirus
Structure determination
Virus structure

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10.1016/j.virol.2010.03.028

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@article{285344f43d2d44d19c07e519254bbe38,

title = "Crystallization and preliminary X-ray diffraction analysis of human adenovirus",

abstract = "Replication-defective and conditionally replicating adenovirus (AdV) vectors are currently being utilized in ~25% of human gene transfer clinical trials. Unfortunately, progress in vector development has been hindered by a lack of accurate structural information. Here we describe the crystallization and preliminary X-ray diffraction analysis of a HAdV5 vector that displays a short flexible fiber derived from HAdV35. Crystals of Ad35F were grown in 100mM HEPES pH 7.0, 200mM Ca(OAc)2, 14% PEG 550 MME, 15% glycerol in 100mM Tris-HCl 8.5. Freshly grown crystals diffracted well to 4.5{\AA} resolution and weakly to 3.5{\AA} at synchrotron sources. HAdV crystals belong to space group P1 with unit cell parameters a=854.03{\AA}, b=855.17{\AA}, c=865.24{\AA}, α=119.57°, Β=91.71°, γ=118.08° with a single particle in the unit cell. Self-rotation and locked-rotation function analysis allowed the determination of the particle orientation. Molecular replacement, density modification and phase-extension procedures are being employed for structure determination.",

keywords = "Capsid structure, Crystallization, Human adenovirus, Structure determination, Virus structure",

author = "Reddy, {V. S.} and Natchiar, {S. K.} and L. Gritton and Mullen, {T. M.} and Stewart, {P. L.} and Nemerow, {G. R.}",

note = "Funding Information: The authors thank Professor Jack Johnson of TSRI for helpful discussions on aspects of crystallization and data collection. The authors also acknowledge Dr. Sangita Venkataraman for her help with the crystallization setups of HAdV, Dr. Irimpan Mathews of SSRL for his initial help with the data collection and Drs. Wladek Minor and Iwona Minor for generously providing the larger version of SCALEPACK program. We also gratefully acknowledge Drs. Robert Fischetti, Nukri Sanishvili and other members at GM/CA CAT for helpful discussions and technical support. GM/CA CAT has been funded in whole or in part with Federal funds from the National Cancer Institute ( Y1-CO-1020 ) and the National Institute of General Medical Science ( Y1-GM-1104 ). The use of the Advance Photon Source was supported by the U.S. Department of Energy, Basic Energy Sciences, Office of Science (contract No. DE-AC02-06CH11357). Funding Information: This work was supported by NIH grants R01 AI070771 to V.S.R and EY011431 and HL54352 to GRN. This is manuscript #20482 at The Scripps Research Institute.",

year = "2010",

month = jun,

doi = "10.1016/j.virol.2010.03.028",

language = "English (US)",

volume = "402",

pages = "209--214",

journal = "Virology",

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T1 - Crystallization and preliminary X-ray diffraction analysis of human adenovirus

AU - Reddy, V. S.

AU - Natchiar, S. K.

AU - Gritton, L.

AU - Mullen, T. M.

AU - Stewart, P. L.

AU - Nemerow, G. R.

N1 - Funding Information: The authors thank Professor Jack Johnson of TSRI for helpful discussions on aspects of crystallization and data collection. The authors also acknowledge Dr. Sangita Venkataraman for her help with the crystallization setups of HAdV, Dr. Irimpan Mathews of SSRL for his initial help with the data collection and Drs. Wladek Minor and Iwona Minor for generously providing the larger version of SCALEPACK program. We also gratefully acknowledge Drs. Robert Fischetti, Nukri Sanishvili and other members at GM/CA CAT for helpful discussions and technical support. GM/CA CAT has been funded in whole or in part with Federal funds from the National Cancer Institute ( Y1-CO-1020 ) and the National Institute of General Medical Science ( Y1-GM-1104 ). The use of the Advance Photon Source was supported by the U.S. Department of Energy, Basic Energy Sciences, Office of Science (contract No. DE-AC02-06CH11357). Funding Information: This work was supported by NIH grants R01 AI070771 to V.S.R and EY011431 and HL54352 to GRN. This is manuscript #20482 at The Scripps Research Institute.

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AB - Replication-defective and conditionally replicating adenovirus (AdV) vectors are currently being utilized in ~25% of human gene transfer clinical trials. Unfortunately, progress in vector development has been hindered by a lack of accurate structural information. Here we describe the crystallization and preliminary X-ray diffraction analysis of a HAdV5 vector that displays a short flexible fiber derived from HAdV35. Crystals of Ad35F were grown in 100mM HEPES pH 7.0, 200mM Ca(OAc)2, 14% PEG 550 MME, 15% glycerol in 100mM Tris-HCl 8.5. Freshly grown crystals diffracted well to 4.5Å resolution and weakly to 3.5Å at synchrotron sources. HAdV crystals belong to space group P1 with unit cell parameters a=854.03Å, b=855.17Å, c=865.24Å, α=119.57°, Β=91.71°, γ=118.08° with a single particle in the unit cell. Self-rotation and locked-rotation function analysis allowed the determination of the particle orientation. Molecular replacement, density modification and phase-extension procedures are being employed for structure determination.

KW - Capsid structure

KW - Crystallization

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KW - Structure determination

KW - Virus structure

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Crystallization and preliminary X-ray diffraction analysis of human adenovirus

Abstract

Bibliographical note

Keywords

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