Large scale purification and refolding of HIV-1 protease from Escherichia coli inclusion bodies. NLM AIDSLINE Important note: Information in this article was accurate in 1994. The state of the art may have changed since the publication date.

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Large scale purification and refolding of HIV-1 protease from Escherichia coli inclusion bodies.

J Protein Chem. 1993 Jun;12(3):323-7. Unique Identifier : AIDSLINE MED/94000339
Hui JO; Tomasselli AG; Reardon IM; Lull JM; Brunner DP; Tomich CS; Heinrikson RL; Upjohn Company, Kalamazoo, Michigan 49001.

Abstract: The protease encoded by the human immunodeficiency virus type 1 (HIV-1) was engineered in Escherichia coli as a construct in which the natural 99-residue polypeptide was preceded by an NH2-terminal methionine initiator. Inclusion bodies harboring the recombinant HIV-1 protease were dissolved in 50% acetic acid and the solution was subjected to gel filtration on a column of Sephadex G-75. The protein, eluted in the second of two peaks, migrated in SDS-PAGE as a single sharp band of M(r) approximately 10,000. The purified HIV-1 protease was refolded into an active enzyme by diluting a solution of the protein in 50% acetic acid with 25 volumes of buffer at pH 5.5. This method of purification, which has also been applied to the purification of HIV-2 protease, provides a single-step procedure to produce 100 mg quantities of fully active enzyme.
Keywords: Acetic Acids Amino Acid Sequence Escherichia coli/*ENZYMOLOGY HIV Protease/CHEMISTRY/*ISOLATION & PURIF HIV-1/ENZYMOLOGY Inclusion Bodies/*ENZYMOLOGY Molecular Sequence Data Protein Folding Recombinant Proteins/CHEMISTRY/ISOLATION & PURIF Viral Proteins/CHEMISTRY/*ISOLATION & PURIF JOURNAL ARTICLEKWDaceticacidsaminoacidsequenceescherichiacoli/KWDenzymologyhivprotease/chemistry/KWDisolation&purifhiv-1/enzymologyinclusionbodies/KWDenzymologymolecularsequencedataproteinfoldingrecombinantproteins/chemistry/isolation&purifviralproteins/chemistry/KWDisolation&purifjournalarticle
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