HIV-2 Protease

Introduction to the Molecule

Retroviral protease from HIV-2 virus is an enzyme important in the life cycle of the virus. It is expressed in the infected cells as a part of Gag-Pol polyprotein from which it is autocatalytycaly released after formation of immature viral particle. The enzyme subsequently cleaves the other parts of viral polyproteins causing the maturation of the virus. In HIV-infected patients the enzyme is a subject of intensive mutagenesis and mutants resistant to applied medcines are produced as a consequence of the selection pressure. HIV-2 protease is active as a homodimer.

Research topic

Others

Description

Total 99 AA. MW: 10.7 kDa (monomer), protein active as dimer

Amino Acid Sequence

Source

E.coli, refolded from inclusion bodies

Purity

Purity as determined by densitometric image analysis: >95%

SDS-PAGE gel

14% SDS-PAGE separation of Human HIV-2 Protease M.W. marker – 10, 20, 30, 40, 60, 80 kDa reduced and heated sample, 2.5 μg/lane

Formulation

20 mM Tris, 20 mM MES, 200 mM NaCl, 10% glycerol, 1 mM EDTA, 0.5 mM DTT, 0.05% PEG 8000, pH 7.0 – filtered (0.4 μm), frozen

Reconstitution

Defrost at ambient temperature.

Storage, Stability/Shelf Life

Store protein at –80°C. Protein remains stable until the expiry date when stored at –80°C. Avoid repeated freezing/thawing cycles.

Quality Control Test

SDS PAGE to determine purity of the protein.

Active site titration by tightly binding inhibitor.

Applications

Crystallography, Inhibitor screening, Kinetic studies

Note

K_m = 740 μM

K_cat = 3 s^-1

K_cat /K_m = 4.1 mM^-1 s^-1 with peptide substrate ATLNFPISPW

Manufactured by AscoProt Biotech

References

• Anderson PC, Lamarre D, Do F, Pargellis C, Pav S, Tong L. Crystal structure of human immunodeficiency virus (HIV) type 2 protease in complex with a reduced amide inhibitor and comparison with HIV-1 protease structures. Proc Natl Acad Sci U S A. 1993 Sep 15;90 (18):8387-91
• Craik CS, Salto R, Rose JR. Regulation of autoproteolysis of the HIV-1 and HIV-2 proteases with engineered amino acid substitutions. J Biol Chem. 1993 Jun 5;268 (16):11939-45
• Weber IT, Ghosh AK, Aniana A, Louis JM, Kovalevsky AY. Structural evidence for effectiveness of darunavir and two related antiviral inhibitors against HIV-2 protease. J Mol Biol. 2008 Dec 5;384 (1):178-92

By research topic:

• Others

By molecule:

• HIV-2 Protease