Inhibition of Viral Fusion Membrane Protein-

Activity via Protein-Specific Protease as the

Next Possible Drug Candidate Against HIV
Infection
“HIV/AIDS: The Greatest Challenge of Our Generation”
Kofi Annan
 By: Mahshid Khatanifar
Acquired immunodeficiency syndrome (AIDS) is the name given to end-stage

disease caused by infection with human immunodeficiency virus (HIV). By almost any

criteria HIV qualifies as one of the world’s deadliest diseases that have ever faced the

humanity. This dreaded infectious disease has claimed the lives of over 25 million people

worldwide and infected 40 million more. In the United States alone, 1.2 million are

infected with the HIV virus and more than 500,000 have died. No virus has been as well

studied or understood as HIV, and yet we are far from controlling this pandemic.

According to the United Nations by the year 2015, in the 60 countries most

affected by AIDS, the total population will be 115 million less than it would be in the

absence of AIDS. Another set of statistics published by UNAIDS/WHO estimated that

AIDS claimed 320,000 lives in 2005 and more than 800 every day in Africa1. Other than

the fact that HIV sacrifices millions and millions of lives every year, it also has a major

impact on social, political, cultural, and economical consequences on human generations.

Therefore for the past 25 years, the pressure caused by all these mentioned factors have

forced the large majortiy of individuals in scientific world to unite against fighting this

deadly virus. Farthermore any attempt for trying to understand this complex diseas will

bring us a step closer to a save a life of a human being.

There are two majore strain of HIV. The new strain of HIV (HIV-2) discovered

off the West coast of Africa is distinctly different than the original HIV-1 strain, while

they have almost the same set of genes and very similar pathological effects. HIV virus

belong to a family of Retroviridae, as well as a member of the Lentivirus genus. HIV-1

exhibits all of the structural characteristics of a typical Lentivirus Retroviridae. It was

originally named a retrovirus because of its particle associated reverse transcriptase, a

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hallmark of the Retroviridae family. It is a 100 to 120 µm enveloped virion with 2 single

stranded RNA.

AIDS is characterized by having an unusually long period from infection with

HIV to the development of symptoms. This period is passed with no apparent symptoms

(asymptomatic carriers) and it seems that the virus is in a state of hibernation. But, in fact

the virus is actively reproduced every day and is in fight with the anti-HIV immune

response. Passing this non-symptomatic period, which may last as long as several

decades now, failure in the immune system will occur resulting in AIDS symptoms.

The HIV-1 virion uses the host cell membrane to form the viral envelope. This envelope

is covered by gp41 and gp120 surface glycoproteins, as well as Major histocompatability

complex class II (MHC II) proteins inserted into the lipid envelope. Inside the lipid

envelope, the matrix formed by Gag protein p17 holding the RNA containing core in

place. The cylindrical core not only stores the viral RNA and various proteins, it also

contains complementary RNA synthesized by the viral reverse transcriptase.

HIV-1 enters the blood stream most commonly through genital or colonic mucosa

during sexual intercourse or other blood to blood contact. The virus enters the cell using

the viral glycoprotein gp120 to interact with the host cellular receptor CD4 and

chemokine receptors CCR5 (predominantly expressed on dendritic cells, macrophages,

and CD4 T cells) and or CXCR4 (expressed on activated T cells), which act as the co-

receptors in HIV’s direct cell fusion-type of infection.

In order for the HIV virus to fuse with the host cell membrane several

conformational changes must occur. The core or the active binding site of the gp120 will

interact and bind a portion of CD4 receptor protein. The core of the gp120 contains the

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bridging beta sheets which connect the inner domain to the outer domain of the receptor.

The CD4 will bind in the active site of gp120 at the junction of bridging sheet, inner

domain and outer domain. This receptor binding with high affinity is mediated by gp120

glycoprotein complex which is recognized by CD4 receptors on lymphocytes. At this

point a second cell surface chemokine receptor interacts with gp120 and form a

complex7.

This interaction releases the HIV fusion protein from gp120 liberating the

previously buried hydrophobic fusion peptide, to insert itself into the plasma membrane.

The fusion protein is a trimer which becomes anchored as an integral membrane protein

in two opposite membranes. The fusion protein then spontaneously rearranges, collapsing

into a tightly packed six-helix bundle. The energy released by this conformational change

is used to pull the two membranes together. This will allow this process to overcome the

high activation energy barrier that normally prevents membrane fusion. As the two

plasma membranes are pulled together the mechanical force created by the helix bundle

will force the water out from the interface and allow the lipids of the two interacting

leaflets of the bilayer to flow between the membranes and form a connection4 (fig 1).

With the basic biology provided designing a drug that could inhibit the virus entry

into the cell at the cell membrane level may seem disputably possible. Therefore the type

of drug that I thought it maybe a possible candidate for anti-HIV treatment will be a

therapeutic, protease type drug that will inhibit the activity of fusion proteins by cleaving

it and disassemble the entire complex, which will farther eliminate the viral entry into the

host cell. To design this drug, identification of the amino acid sequence of the viral

genome that will encode for the fusion peptide in viral genome is necessary. After

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cloning that specific segment of the genome in E-Coli vector and translating into the

corresponding proteins (HIV fusion peptide), with the help of the crystal structure of the

peptide and some computer analyzing programs such as QSAR , we could then choose a

protease that is able to cleave the peptide bond from the N terminus and lead to

disassembly of the helix bundle, which will then deactivate the fusion protein and farther

inhibit the viral entry.

fig. 1. the red hairpin structure is part of the fusion peptide which will promote complete
fusions of both host and viral membranes via the mechanism explained above.

Experiments have shown that the ability of the HIV-1 virus to infect cells can be

greatly diminished by deactivation of the N-terminal fusion peptide of its glycoprotein

gp413. Other studies also shown that the fusion domain HIV-1 envelope glycoprotein

(gp120-gp41) includes a conserved hydrophobic region that is located at the N terminus

of the gp41, and as a result a V2E mutant of this region has been shown to dominantly

interfere with wild-type envelope-mediated syncytium formation and virus infectivity6. In

recent studies using the same basic biology, researchers have developed molecules such

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as NSC 13778 that are designed to block the entry of X4-, R5-, and X4/R5-tropic HIV-1

strains into CD4+ cells2.

According to FDA the guidelines to approve a certain drug is estimated to be

between 9.75 years to 7 years or 5.5 years for drugs treating serious diseases (probably

such as HIV). Although this data is only true when the drug is successful through each

phase of the FDA designed procedures8. The most important factor in developing a

biomedical product and in this case an anti-HIV drug is to be able to understand the

“tricks” that the virus uses and apply that information, to produce methods for inhibiting

the certain pathways. The other thing is to keep in mind that the ideal drug will be the

one that prevents an infection before the virus enters the cell. Therefore inhibiting the

fusion peptide activity may hold a promising future in anti-HIV drug development.

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Reference
1.
http://data.unaids.org/pub/GlobalReport/2006/2006_GR_CH04_en.pdf

2.
Yang, Q-e., Stephen, AG., Adelsberger, WJ., Roberts, P., Zhu, W., Currens, M., Feng,
YX., Crise, BJ., Gorelick, RJ., Rein, AR., Fisher, RJ., Shoemaker, RH., Sei, S. Discovery
of Small-Molecule HIV-1 Entry Inhibitors that Target the gp120-binding domain of
CD4. Journal of Virology. 2005; 79:6122–6133.

3.
Maddox, MW., Longo, ML. Conformational partitioning of the fusion peptide of HIV-
1 gp41 and its structural analogs in bilayer membranes. Biophysical Journal .
December 2002.

4.
Sackett, K., Wexler-Cohen, Y., Shai, Y. Characterization of the HIV N-terminal
Fusion Peptide-containing Region in Context of Key gp41 Fusion Conformations.
Journal of Biological Chemistry. August 4, 2006: 281, Issue 31, 21755-21762.

5.
http://www.callutheran.edu/Academic_Programs/Departments/BioDev/omm/jmol/hiv_g
p120/gp120.html

6.
Kliger Y, Aharoni A, Rapaport D, Jones P, Blumenthal R, Shai Y. Fusion peptides
derived from the HIV type 1 glycoprotein 41 associate within phospholipid membranes
and inhibit cell-cell Fusion. Structure-function study. Journal of Biological Chemistry.
1997 May 23; 272(21):13496-505.

7.
Immunobiology, the Immune System in Health and Disease, 6th Ed. by C. Janeway et. al.

8.
www.FDA.gov