The Refined Crystal Structure of Hexon, the Major Coat Protein of Adenovirus Type 2, at 2·9 Å Resolution

Athappilly, F.; Murali, Ramachandran; Rux, John J.; Cai, Zhaoping; Burnett, Roger M.

doi:10.1006/jmbi.1994.1593

Cited by 122 publications

(81 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This assumption is supported by the presence of a stretch of acidic residues in the sequence of the Ad2 and Ad5 hexons (position 133 to 161) which is partially exposed on the outer surface of the particle. 7 Interestingly, the interaction with the adsorbent is not mediated by these acidic residues only since Ad3, Ad4 and Ad7 which do not contain this sequence of acidic residues are also retained on this adsorbent (Table 6). However, a trend was found between the charge of the hexon L1 loop (which contains the region 133 to 166) and the HPLC retention time (Table 6), which indicates that the acidic region of the hexon protein is very likely to be a major determinant of the strong interaction with Q Sepharose XL observed with Ad2 and Ad5.…”

Section: Discussionmentioning

confidence: 99%

“…It leads to the generation of AV 1.0 CMV.lacZ adenovirus. Plasmid pXL3497 derives from pXL3185 and has a sequence inserted at position 32784 (Ad5 coordinates) which corresponds to the addition of a (Gly-Ser) 5 -(Lys) 7 peptide at the C-terminus of the fiber protein. 20 The corresponding adenovirus, AV 1.k CMV.lacZ, was described as AdZ.F(pK7)␤gal by Wickham et al 20 Plasmid pXL3615 is an RK2-derived vector containing an Ad5 genome carrying deletions in the E1 region (from position 386 to position 3512), in the E3 region (from position 28 592 to position 30 470) and in the E4 region (from position 33 423 to position 35 356).…”

Section: Adenoviral Genomementioning

confidence: 99%

See 1 more Smart Citation

An improved anion-exchange HPLC method for the detection and purification of adenoviral particles

Blanche¹,

Cameron²,

Barbot³

et al. 2000

Gene Ther

View full text Add to dashboard Cite

We have developed an anion-exchange high-performance liquid chromatography (HPLC) method using Q Sepharose XL (Amersham Pharmacia Biotech) as adsorbent to analyze samples containing adenovirus. This method has several major advantages over the HPLC method previously described for quantitating particles, namely (1) a Ͼ10-fold improvement in the detection limit of adenovirus in crude preparations; (2) absence of interferences originating from nucleic acids and proteins which usually contaminate crude samples; (3) unprecedented sharpness and symmetry of adenovirus peak, rendering the identification of the viral peak unambiguous, even in extremely crude and dilute prep-

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Adenoviral Genomementioning

confidence: 99%

An improved anion-exchange HPLC method for the detection and purification of adenoviral particles

Blanche¹,

Cameron²,

Barbot³

et al. 2000

Gene Ther

View full text Add to dashboard Cite

show abstract

“…7,8 Even more detail on the protein level has been obtained from the crystal structures of hexon and the fiber knob. 9,10 These studies have been crucial to coat protein engineering efforts because they have taken the guesswork out of where to insert ligands into the coat proteins. In addition, the structure of the Ad5 knob has been instrumental for our group in delineating the binding site for CAR on the fiber.…”

Section: Appreciating the Biologymentioning

confidence: 99%

Targeting adenovirus

2000

View full text Add to dashboard Cite

“…1A) has more precisely defined the location of the surface loops than a homology model. In combination with the crystal structures of Ad2 (1,25) and Ad5 (24,25) hexons, the new structure has given an improved sequence alignment for all hexons (J. J. Rux and R. M. Burnett, unpublished). Nonetheless, the specific locations of neutralizing epitopes have not been previously identified for any adenovirus.…”

mentioning

confidence: 99%

Structure-Based Identification of a Major Neutralizing Site in an Adenovirus Hexon

Pichla-Gollon

Drinker

Zhou

et al. 2007

J Virol

Self Cite

View full text Add to dashboard Cite

Virus-specific neutralizing antibodies present an obstacle to the effective use of adenovirus vectors for gene therapy and vaccination. The specific sites recognized by neutralizing antibodies have not been identified for any adenovirus, but they have been proposed to reside within the hexon, in small regions of the molecule that are exposed on the capsid surface and possess sequences that vary among serotypes. We have mapped the epitopes recognized by a panel of seven hexon-specific monoclonal antibodies that neutralize the chimpanzee adenovirus 68 (AdC68). Surface plasmon resonance experiments revealed that the antibodies compete for a single hexon binding site, and experiments with synthetic peptides indicated that this site resides within just one small surface loop. Mutations within this loop (but not in other surface loops) permitted virus to escape neutralization by all seven monoclonal antibodies and to resist neutralization by polyclonal antisera obtained from animals immunized against AdC68. These results indicate that a single small surface loop defines a major neutralization site for AdC68 hexon.Modified adenoviruses have been widely used as vehicles for gene delivery and as vaccine vectors. Most adenovirus vectors in use have been derived from the human serotype 5 (Ad5). As almost all human adults have been exposed to Ad5, they possess neutralizing antibodies to Ad5 that limit the efficiency of the virus as a delivery vector (7,33,42). Approaches to circumvent the problem of preexisting immunity include chemical modification of Ad5 surface proteins to mask the neutralizing epitopes (4,19,29) and replacement of the immunogenic capsid proteins with those of other serotypes (21,23,32,41,44). Many investigators are also exploring the use of rare human serotypes (such as human Ad48) or nonhuman adenoviruses (including those derived from dogs, fowl, and nonhuman primates) to which humans are not usually immune (2, 6, 13, 16-18, 22, 30).An alternative approach is to identify the specific sites on adenovirus that are recognized by neutralizing antibodies and then modify those sites to generate mutants capable of escaping neutralization. One nonhuman serotype that has been proposed as an alternative vector for vaccination is the chimpanzee adenovirus 68 (AdC68) (42). AdC68 is not neutralized by most human adult sera and elicits a strong transgene productspecific immune response in animals already immune to Ad5 (7,33,42). However, because one immunization with an AdC68 vector will induce serotype-specific immunity, multipledose immunization regimens may require the availability of additional vectors. Production of antigenically modified vectors would be facilitated if the epitopes recognized by the neutralizing antibodies were well characterized.The adenovirus capsid is an icosahedron with long fibers projecting from the vertices. Twelve copies of the trimeric major capsid protein, hexon, form each of the 20 triangular facets of the icosahedron; trimeric fibers are inserted into the pentameric penton bases at...

show abstract

The Refined Crystal Structure of Hexon, the Major Coat Protein of Adenovirus Type 2, at 2·9 Å Resolution

Cited by 122 publications

References 0 publications

An improved anion-exchange HPLC method for the detection and purification of adenoviral particles

An improved anion-exchange HPLC method for the detection and purification of adenoviral particles

Targeting adenovirus

Structure-Based Identification of a Major Neutralizing Site in an Adenovirus Hexon

Contact Info

Product

Resources

About