EurekaMag
+ Translate
+ Most Popular
The pigeon tick (Argas reflexus): its biology, ecology, and epidemiological aspects
Prevalence of hemoglobin abnormalities in Kebili (Tunisian South)
Lipogranuloma: a preventable complication of dacryocystorhinostomy
Value of basal plasma cortisol assays in the assessment of pituitary-adrenal insufficiency
Bees from the Belgian Congo. The acraensis group of Anthophora
Placing gingival retraction cord
Total serum IgE, allergy skin testing, and the radioallergosorbent test for the diagnosis of allergy in asthmatic children
Acariens plumicoles Analgesoidea parasites des oiseaux du Maroc
Injuries of terminal phalanges of the fingers in children
Biology of flowering and nectar production in pear (Pyrus communis)
Das Reliktvorkommen der Aspisviper (Vipera aspis L.) im Schwarzwald
Hydrological modelling of drained blanket peatland
Pathologic morphology and clinical significance of the anomalous origin of the left circumflex coronary artery from the right coronary artery. General review and autopsy analysis of 30 cases
Cyto genetic analyses of lymphocyte cultures after exposure to calcium cyclamate
Axelrodia riesei, a new characoid fish from Upper Rio Meta in Colombia With remarks concerning the genus Axelrodia and description of a similar, sympatric, Hyphessobrycon-species
Favorable evolution of a case of tuberculosis of pancreas under antibiotic action
RIFM fragrance ingredient safety assessment, Valencene, CAS Registry Number 4630-07-3
Parenteral microemulsions: an overview
Temperate pasture: management for grazing and conservation
Evaluation of a new coprocessed compound based on lactose and maize starch for tablet formulation
Thermal expansion and cracking of three confined water-saturated igneous rocks to 800C
Revision of the genera of the tribe Stigmoderini (Coleoptera: Buprestidae) a discussion of phylogenetic relationships
Anal tuberculosis. Report of a case
Gastric tuberculosis in the past and present
Adaptive responses of the cardiovascular system to prolonged spaceflight conditions: assessment with Holter monitoring

High level expression of 5-helix protein in HIV gp41 heptad repeat regions and its virus fusion-inhibiting activity


High level expression of 5-helix protein in HIV gp41 heptad repeat regions and its virus fusion-inhibiting activity



Sheng Wu Gong Cheng Xue Bao 25(3): 435-440



ISSN/ISBN: 1000-3061

PMID: 19621587

The artificial 5-helix can inhibit the formation of trimer-of-hairpins structure during the course of HIV-directed membrane fusion and then inhibit human immunodeficiency virus (HIV) infecting target cells. But 5-helix was apt to form inclusion body when expressed directly in prokaryotic cell and was difficult to renature, which causes inconvenience to future study. We found a proper expression vector by simulating protein structure. We simulated its proper conformation in two vectors pGEX-6P-1 and pET44b by homology modeling. The contrast of conformations showed that the energy of salvation of its fusion protein with NusA in vector pET44b was higher than its fusion protein with glutathione-S-transferase (GST) in pGEX-6P-1 and its restriction site lay on the surface of its fusion protein in vector pET44b. 5-helix gene was amplified from pGEX-6P-1-5H by PCR, and was ligated to pET44b to construct recombinant vector pET44b-PSP-5Helix after tested correctly by enzymes digestion. The recombinant vector was transformed into Escherichia coli BL21 (DE3) to express 5-helix protein at different temperatures. Aim protein was purified with Ni column and GST column, and was determined by SDS-PAGE. Then the purified 5 -Helix was used to test the inhibitive activity of pseudo HIV virus infecting GHOST-CXCR4. Results show that its fusion protein with NusA can be effectively soluble expressed and easier to be cleaved, and that the purified 5-helix can efficiently inhibit pseudo HIV virus infecting GHOST-CXCR4 and its IC50 value is (22.77 +/- 5.64) nmol/L, which lay the foundation to further discuss the application in HIV-1 infection.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 053536359

Download citation: RISBibTeXText

Related references

Design and analysis of post-fusion 6-helix bundle of heptad repeat regions from Newcastle disease virus F protein. Protein Engineering 16(5): 373-379, 2003

Cloning and expression of heptad repeat regions of the Newcastle disease virus fusion protein. Sheng Wu Gong Cheng Xue Bao 17(6): 631-634, 2001

Mutations in the fusion peptide and heptad repeat regions of the Newcastle disease virus fusion protein block fusion. Journal of Virology 68(11): 7654-7658, 1994

Six-helix bundle assembly and characterization of heptad repeat regions from the F protein of Newcastle disease virus. Journal of General Virology 83(Pt 3): 623-629, 2002

Interaction of peptides with sequences from the Newcastle disease virus fusion protein heptad repeat regions. Journal of Virology 73(7): 5945-5956, 1999

A Heptad Repeat in Herpes Simplex Virus 1 gH, Located Downstream of the a-Helix with Attributes of a Fusion Peptide, Is Critical for Virus Entry and Fusion. Journal of Virology 79(11): 42-9, 2005

A Heptad Repeat in Herpes Simplex Virus 1 gH, Located Downstream of the -Helix with Attributes of a Fusion Peptide, Is Critical for Virus Entry and Fusion. Journal of Virology 81(5): 2540-2540, 2007

A heptad repeat in herpes simplex virus 1 gH, located downstream of the alpha-helix with attributes of a fusion peptide, is critical for virus entry and fusion. Journal of Virology 79(11): 7042-7049, 2005

The antibodies directed against N-terminal heptad-repeat peptide of hRSV fusion protein and its analog-5-Helix inhibit virus infection in vitro. Biochemical and Biophysical Research Communications 331(4): 1358-1364, 2005

A heptad repeat in herpes simplex virus 1 gH, located downstream of the alpha-helix with attributes of a fusion peptide, is critical for virus entry and fusion (vol 79, pg 7042, 2005). Journal of Virology 81(5): 2540, 2007

Mutations in the fusion peptide and adjacent heptad repeat inhibit folding or activity of the Newcastle disease virus fusion protein. Journal of Virology 75(17): 7934-7943, 2001

Biochemical and biophysical analysis of heptad repeat regions from the fusion protein of Menangle virus, a newly emergent paramyxovirus. Archives of Virology 148(7): 1301-1316, 2003

Heptad repeat 2 in herpes simplex virus 1 gH interacts with heptad repeat 1 and is critical for virus entry and fusion (vol 80, pg 2216, 2006). Journal of Virology 81(5): 2541, 2007

Interaction between heptad repeat 1 and 2 regions in spike protein of SARS-associated coronavirus: implications for virus fusogenic mechanism and identification of fusion inhibitors. Lancet 363(9413): 938-947, 2004

Heptad Repeat 2 in Herpes Simplex Virus 1 gH Interacts with Heptad Repeat 1 and Is Critical for Virus Entry and Fusion. Journal of Virology 81(5): 2541-2541, 2007