Design and characterization of an engineered gp41 protein from human immunodeficiency virus-1 as a tool for drug discovery.
Two new proteins of approximately 70 amino acids in length, corresponding to an unnaturally-linked N- and C-helix of the ectodomain of the gp41 protein from the human immunodeficiency virus (HIV) type 1, were designed and characterized. A designed tripeptide links the C-terminus of the C-helix with the N-terminus of the N-helix in a circular permutation so that the C-helix precedes the N-helix in sequence. In addition to the artificial peptide linkage, the C-helix is truncated at its N-terminus to expose a region of the N-helix known as the "Trp-Trp-Ile" binding pocket. Sedimentation, crystallographic, and nuclear magnetic resonance studies confirmed that the protein had the desired trimeric structure with an unoccupied binding site. Spectroscopic and centrifugation studies demonstrated that the engineered protein had ligand binding characteristics similar to previously reported constructs. Unlike previous constructs which expose additional, shallow, non-conserved, and undesired binding pockets, only the single deep and conserved Trp-Trp-Ile pocket is exposed in the proteins of this study. This engineered version of gp41 protein will be potentially useful in research programs aimed at discovery of new drugs for therapy of HIV-infection in humans.
Stewart
Kent D
KD
Department of Structural Biology, Abbott Laboratories, Building AP10, 100 Abbott Park Road, Abbott Park, IL 60064, USA. kent.stewart@abbott.com
Steffy
Kevin
K
Harris
Kevin
K
Harlan
John E
JE
Stoll
Vincent S
VS
Huth
Jeffrey R
JR
Walter
Karl A
KA
Gramling-Evans
Emily
E
Mendoza
Renaldo R
RR
Severin
Jean M
JM
Richardson
Paul L
PL
Barrett
Leo W
LW
Matayoshi
Edmund D
ED
Swift
Kerry M
KM
Betz
Stephen F
SF
Muchmore
Steve W
SW
Kempf
Dale J
DJ
Molla
Akhter
A
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D015195:Drug Design; D015700:HIV Envelope Protein gp41 / Q000737:chemistry* / Q000235:genetics; D015497:HIV-1 / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D011487:Protein Conformation; D015202:Protein Engineering
D015700:HIV Envelope Protein gp41