Three-dimensional structure of human cytomegalovirus protease

During assembly of herpesvirus capsids, a protein scaffold self-assembles to ring-like structures forming the scaffold of the spherical procapsids. Proteolytic activity of the herpesvirus maturational protease causes structural changes that result in angularization of the capsids. In those mature icosahedral capsids, the packaging of viral DNA into the capsids can take place. The strictly regulated protease is called assemblin. It is inactive in its monomeric state and activated by dimerization. The structures of the dimeric forms of several assemblins from all herpesvirus subfamilies have been elucidated in the last two decades. They revealed a unique serine-protease fold with a catalytic triad consisting of a serine and two histidines. Inhibitors that disturb dimerization by binding to the dimerization area were found recently. Additionally, the structure of the monomeric form of assemblin from pseudorabies virus and some monomer-like structures of Kaposi's sarcoma-associated herpesvirus assemblin were solved. These findings are the proof-of-principle for the development of new anti-herpesvirus drugs. Therefore, the most important information on this fascinating and unique class of proteases is summarized here.

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“…2) [58,59]. The stabilization of the oxyanion hole is realized in a similar manner in all serine proteases as well.…”

Section: Assemblinsmentioning

confidence: 70%

“…The first crystal structures of an assemblin were published by three groups in 1996 [58,59,83]. All three structures are of dimeric HCMV-assemblin.…”

Section: Structurementioning

confidence: 99%

Assemblins as maturational proteases in herpesviruses

Zühlsdorf

Hinrichs

2017

Journal of General Virology

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“…The amino-acid sequences of herpesvirus proteases have no detectable homology to other serine proteases or hydrolases. The crystal structure of HCMV protease has recently been determined in our laboratory by the seleno-methionyl multiple-wavelength anomalous diffraction (MAD) technique (Tong et al, 1996), and in other laboratories by the heavy-atom isomorphous replacement technique (Qiu et al, 1996;Shieh et al, 1996;Chen et al, 1996). It showed that HCMV protease possesses a new polypeptide backbone fold; confirmed that Ser132 and His63 are in close proximity in space; and identified His157 as the possible third member of the catalytic triad.…”

Section: Introductionmentioning

confidence: 95%

Experiences from the Structure Determination of Human Cytomegalovirus Protease

Tong

Qian²,

Davidson³

et al. 1997

Acta Cryst D

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Several obstacles were encountered and overcome during the structure determination of human cytomegalovirus protease. Dehydration of crystals, by exposing them to higher concentrations of the precipitant, reduced the mosaicity of the crystals and may have also resolved their microscopic twinning. The initial phase information was obtained with the selenomethionyl multiple-wavelength anomalous diffraction technique. However, site-specific mutagenesis was required to introduce extra Met residues into the protease. The phase information had to be improved by non-crystallographic symmetry averaging, initially among three 'crystal forms'. A change in the composition of the artificial mother liquor led to a significant improvement, from 3.0 and 2.0A resolution, in the diffraction quality of the crystals. The experiences reported here may prove useful to structure determination of other proteins.

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“…They exist in a monomer-dimer equilibrium in vitro where only the dimer is active [5][6][7]. Xray crystallographic studies on the protease from CMV, HSV1, HSV2, VZV, KSHV and EBV have shown the presence of a conserved dimer structure [8][9][10][11][12][13][14].…”

Section: Introductionmentioning

confidence: 99%

Kinetics, inhibition and oligomerization of Epstein‐Barr virus protease

Buisson

Rivail

Hernandez³

et al. 2006

FEBS Letters

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Epstein-Barr virus (EBV) is an omnipresent human virus causing infectious mononucleosis and EBV associated cancers. Its protease is a possible target for antiviral therapy. We studied its dimerization and enzyme kinetics with two enzyme assays based either on the release of paranitroaniline or 7-amino-4-methylcoumarin from labeled pentapeptide (Ac-KLVQA) substrates. The protease is in a monomer-dimer equilibrium where only dimers are active. In absence of citrate the K d is 20 lM and drops to 0.2 lM in presence of 0.5 M citrate. Citrate increases additionally the activity of the catalytic sites. The inhibitory constants of different substrate derived peptides and a-keto-amide based inhibitors, which have at best a K i of 4 lM, have also been evaluated.

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Three-dimensional structure of human cytomegalovirus protease

Cited by 156 publications

References 28 publications

Assemblins as maturational proteases in herpesviruses

Assemblins as maturational proteases in herpesviruses

Experiences from the Structure Determination of Human Cytomegalovirus Protease

Kinetics, inhibition and oligomerization of Epstein‐Barr virus protease

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