scholarly journals Contribution of Light Chain Residues to High Affinity Binding in an HIV-1 Antibody Explored by Combinatorial Scanning Mutagenesis

Biochemistry ◽  
2010 ◽  
Vol 49 (26) ◽  
pp. 5464-5472 ◽  
Author(s):  
Gustavo F. Da Silva ◽  
Joseph S. Harrison ◽  
Jonathan R. Lai
Keyword(s):  
Light Chain ◽  
Affinity Binding ◽  
High Affinity Binding ◽  
High Affinity ◽  
Hiv 1

scholarly journals CDK9 Autophosphorylation Regulates High-Affinity Binding of the Human Immunodeficiency Virus Type 1 Tat–P-TEFb Complex to TAR RNA

2000 ◽  
Vol 20 (18) ◽  
pp. 6958-6969 ◽  
Author(s):  
Mitchell E. Garber ◽  
Timothy P. Mayall ◽  
Eric M. Suess ◽  
Jill Meisenhelder ◽  
Nancy E. Thompson ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Affinity Binding ◽  
High Affinity Binding ◽  
High Affinity ◽  
Immunodeficiency Virus ◽  
Tar Rna ◽  
Cdk9 Phosphorylation ◽  
Hiv 1

ABSTRACT Human immunodeficiency virus type 1 (HIV-1) Tat interacts with cyclin T1 (CycT1), a regulatory partner of CDK9 in the positive transcription elongation factor (P-TEFb) complex, and binds cooperatively with CycT1 to TAR RNA to recruit P-TEFb and promote transcription elongation. We show here that Tat also stimulates phosphorylation of affinity-purified core RNA polymerase II and glutathioneS-transferase–C-terminal-domain substrates by CycT1-CDK9, but not CycH-CDK7, in vitro. Interestingly, incubation of recombinant Tat–P-TEFb complexes with ATP enhanced binding to TAR RNA dramatically, and the C-terminal half of CycT1 masked binding of Tat to TAR RNA in the absence of ATP. ATP incubation lead to autophosphorylation of CDK9 at multiple C-terminal Ser and Thr residues, and full-length CycT1 (amino acids 728) [CycT1(1–728)], but not truncated CycT1(1–303), was also phosphorylated by CDK9. P-TEFb complexes containing a catalytically inactive CDK9 mutant (D167N) bound TAR RNA weakly and independently of ATP, as did a C-terminal truncated CDK9 mutant that was catalytically active but unable to undergo autophosphorylation. Analysis of different Tat proteins revealed that the 101-amino-acid SF2 HIV-1 Tat was unable to bind TAR with CycT1(1–303) in the absence of phosphorylated CDK9, whereas unphosphorylated CDK9 strongly blocked binding of HIV-2 Tat to TAR RNA in a manner that was reversed upon autophosphorylation. Replacement of CDK9 phosphorylation sites with negatively charged residues restored binding of CycT1(1–303)-D167N-Tat, and rendered D167N a more potent inhibitor of transcription in vitro. Taken together, these results demonstrate that CDK9 phosphorylation is required for high-affinity binding of Tat–P-TEFb to TAR RNA and that the state of P-TEFb phosphorylation may regulate Tat transactivation in vivo.

scholarly journals A High Affinity Binding Site for the HIV-1 Nucleocapsid Protein

1997 ◽  
Vol 25 (5) ◽  
pp. 1042-1049 ◽  
Author(s):  
J. A. Berglund ◽  
B. Charpentier ◽  
M. Rosbash
Keyword(s):  
Binding Site ◽  
Nucleocapsid Protein ◽  
Affinity Binding ◽  
High Affinity Binding ◽  
High Affinity ◽  
High Affinity Binding Site ◽  
Hiv 1

Design and synthesis of lipid-coupled inositol 1,2,3,4,5,6-hexakisphosphate derivatives exhibiting high-affinity binding for the HIV-1 MA domain

2014 ◽  
Vol 12 (27) ◽  
pp. 5006-5022 ◽  
Author(s):  
Hiroshi Tateishi ◽  
Kensaku Anraku ◽  
Ryoko Koga ◽  
Yoshinari Okamoto ◽  
Mikako Fujita ◽  
...  
Keyword(s):  
Hydrophobic Interactions ◽  
Affinity Binding ◽  
High Affinity Binding ◽  
Design And Synthesis ◽  
High Affinity ◽  
Electrostatic And Hydrophobic Interactions ◽  
Hiv 1

Lipid-coupled inositol 1,2,3,4,5,6-hexakisphosphate binds to HIV-1 MA tightly through both electrostatic and hydrophobic interactions.

Incorporation of a Non-Nucleotide Bridge into Hairpin Oligonucleotides Capable of High-Affinity Binding to the Rev Protein of HIV-1†

Biochemistry ◽  
1996 ◽  
Vol 35 (16) ◽  
pp. 5339-5344 ◽  
Author(s):  
Jeffrey S. Nelson ◽  
Lori Giver ◽  
Andrew D. Ellington ◽  
Robert L. Letsinger
Keyword(s):  
Affinity Binding ◽  
High Affinity Binding ◽  
High Affinity ◽  
Hairpin Oligonucleotides ◽  
Hiv 1 ◽  
Rev Protein

scholarly journals A Specific RNA Structural Motif Mediates High Affinity Binding by the HIV-1 Nucleocapsid Protein (NCp7)

Virology ◽  
1996 ◽  
Vol 225 (2) ◽  
pp. 306-315 ◽  
Author(s):  
Patrick Allen ◽  
Brian Collins ◽  
David Brown ◽  
Zdenek Hostomsky ◽  
Larry Gold
Keyword(s):  
Nucleocapsid Protein ◽  
Structural Motif ◽  
Affinity Binding ◽  
High Affinity Binding ◽  
High Affinity ◽  
Hiv 1

scholarly journals Structural basis for the inhibition of HIV-1 Nef by a high-affinity binding single-domain antibody

Retrovirology ◽  
2014 ◽  
Vol 11 (1) ◽  
pp. 24 ◽  
Author(s):  
Sebastian Lülf ◽  
Julie Matz ◽  
Marie-Christine Rouyez ◽  
Annika Järviluoma ◽  
Kalle Saksela ◽  
...  
Keyword(s):  
Single Domain ◽  
Structural Basis ◽  
Affinity Binding ◽  
Single Domain Antibody ◽  
High Affinity Binding ◽  
High Affinity ◽  
Domain Antibody ◽  
Hiv 1
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