Selection of scFvs specific for HBV DNA polymerase using ribosome display

DNA replication is vital for an organism to proliferate and lying at the heart of this process is the enzyme DNA polymerase. Most DNA polymerases have a similar three dimensional fold, akin to a human right hand, despite differences in sequence homology. This structural homology would predict a relatively unvarying mechanism for DNA synthesis yet various polymerases exhibit markedly different properties on similar substrates, indicative of each type of polymerase being prescribed to a specific role in DNA replication. Several key conformational steps, discrete states, and structural moieties have been identified that contribute to the array of properties the polymerases exhibit. The ability of carcinogenic adducts to interfere with conformational processes by directly interacting with the protein explicates the mutagenic consequences these adducts impose. Recent studies have identified novel states that have been hypothesised to test the fit of the nascent base pair, and have also shown the enzyme to possess a lively quality by continually sampling various conformations. This review focuses on the homologous structural changes that take place in various DNA polymerases, both replicative and those involved in adduct bypass, the role these changes play in selection of a correct substrate, and how the presence of bulky carcinogenic adducts affects these changes.

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Ribosome Display for the Selection of Sac7d Scaffolds

Ribosome Display and Related Technologies - Methods in Molecular Biology ◽

10.1007/978-1-61779-379-0_18 ◽

2011 ◽

pp. 315-331 ◽

Cited By ~ 14

Author(s):

Barbara Mouratou ◽

Ghislaine Béhar ◽

Lauranne Paillard-Laurance ◽

Stéphane Colinet ◽

Frédéric Pecorari

Keyword(s):

Ribosome Display ◽

Selection Of

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In vitro infection of human hepatoma cells (HepG2) with hepatitis B virus (HBV): spontaneous selection of a stable HBV surface antigen-producing HepG2 cell line containing integrated HBV DNA sequences

Journal of General Virology ◽

10.1099/0022-1317-75-10-2681 ◽

1994 ◽

Vol 75 (10) ◽

pp. 2681-2689 ◽

Cited By ~ 18

Author(s):

H. Mabit ◽

S. Dubanchet ◽

F. Capel ◽

C. Dauguet ◽

M.-A. Petit

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Dna Sequences ◽

Hbv Dna ◽

Human Hepatoma ◽

Hepg2 Cell Line ◽

Human Hepatoma Cells ◽

B Virus ◽

Selection Of

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Rapid Selection of High-Affinity Antibody scFv Fragments Using Ribosome Display

Antibody Engineering - Methods in Molecular Biology ◽

10.1007/978-1-4939-8648-4_13 ◽

2018 ◽

pp. 235-268 ◽

Cited By ~ 1

Author(s):

Birgit Dreier ◽

Andreas Plückthun

Keyword(s):

Ribosome Display ◽

High Affinity ◽

Selection Of

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Inhibitory Effects of Acyclic Nucleoside Phosphonate Analogues on Hepatitis B Virus DNA Synthesis in HB611 Cells

Antiviral Chemistry and Chemotherapy ◽

10.1177/095632029400500201 ◽

1994 ◽

Vol 5 (2) ◽

pp. 57-63 ◽

Cited By ~ 23

Author(s):

T. Yokota ◽

K. Konno ◽

S. Shigeta ◽

A. Holy ◽

J. Balzarini ◽

...

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Dna Synthesis ◽

Dna Polymerase ◽

Hbv Dna ◽

Inhibitory Effects ◽

B Virus ◽

Acyclic Nucleoside ◽

Cellular Dna ◽

Acyclic Nucleoside Phosphonate

By using an assay system based on a human hepatoblastoma cell line (HB611) that continuously synthesizes hepatitis B virus (HBV) DNA, 56 acyclic nucleoside phosphonate analogues were examined for their inhibitory effects on HBV DNA synthesis. The following compounds were found to inhibit HBV DNA synthesis at concentrations that were significantly lower than their minimum cytotoxic concentrations; 9-(2-phosphonylmethoxyethyl)adenine (PMEA), 9-(2-phosphonylmethoxyethyl) guanine(PMEG), 9-(2-phosphonylmethoxyethyl) guanine ethyl ester (PMEGEE), 9 - (2 - phosphonylmethoxyethyl) - 1 - deazaadenine (PMEC1A), 9-(2-phosphonylmethoxyethyl)-2,6-diaminopurine (PMEDAP), ( S)-9-(3-hydroxy-2-phosphonylmethoxypropyl)adenine (HPMPA), 9-(3-isopropoxy-2-phosphonylmethoxypropyl)adenine (IPPMPA), 9-( RS)-(2-phosphonylmethoxypropyl)adenine (PMPA) and 9-(3-hydroxy-2-phosphonylmethoxypropyl)-2, 6-diaminopurine (HPMPDAP). The most selective compounds (with indexes greater than 100) were PMEDAP, PMEA, IPPMPA, and PMPA. Acyclic pyrimidine nucleoside phosphonate analogues did not prove markedly selective as anti-HBV agents. Diphosphoryl derivatives of some acyclic purine nucleoside phos-phonates (i.e. PMEA, PMEDAP, HPMPA) were prepared. They proved inhibitory to HBV DNA polymerase but not cellular DNA polymerase α.

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