Enzymatic activity of a synthetic 99 residue protein corresponding to the putative HIV-1 protease

Schneider, Jens; Kent, Stephen B. H.

doi:10.1016/0092-8674(88)90199-7

Cited by 249 publications

(119 citation statements)

References 25 publications

Supporting

Mentioning

114

Contrasting

Unclassified

Order By: Relevance

“…The sequence and synthesis of these peptides have been reported previously (Schneider & Kent, 1988), the only difference here being that the p24/p15 peptide was synthesized with a C-terminal amide rather than free acid. The p17/p24 peptide contains one known HIV-1 protease cleavage site (Tyr-Pro), and the p24/p15 peptide has two cleavage sites (Met-Met and Leu-Ala).…”

Section: Properties Of Btd Hiv-i Proteasementioning

confidence: 99%

“…From a protein engineering perspective, the HIV-1 protease represents an ideal enzyme to study because it can be prepared by total chemical synthesis (Schneider & Kent, 1988). The synthetic enzyme has been prepared in crystalline form and was used to solve the three-dimensional structure of both the native molecule and the enzyme complexed with several dif- ferent substrate-derived inhibitors Swain et al, 1990;Jaskolski et al, 1991).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Structural engineering of the HIV‐1 protease molecule with a β‐turn mimic of fixed geometry

1993

View full text Add to dashboard Cite

An important goal in the de novo design of enzymes is the control of molecular geometry. To this end, an analog of the protease from human immunodeficiency virus 1 (HIV-1 protease) was prepared by total chemical synthesis, containing a constrained, nonpeptidic type 11' 0-turn mimic of predetermined three-dimensional structure.The mimic &turn replaced residues GlyI6,l7 in each subunit of the homodimeric molecule. These residues constitute the central amino acids of two symmetry-related type I' 0-turns in the native, unliganded enzyme. The 0-turn mimic-containing enzyme analog was fully active, possessed the same substrate specificity as the GlyI67l7-containing enzyme, and showed enhanced resistance to thermal inactivation. These results indicate that the precise geometry of the @turn at residues 15-18 in each subunit is not critical for activity, and that replacement of the native sequence with a rigid 0-turn mimic can lead to enhanced protein stability. Finally, the successful incorporation of a fixed element of secondary structure illustrates the potential of a "molecular kit set" approach to protein design and synthesis.

show abstract

Section: Properties Of Btd Hiv-i Proteasementioning

confidence: 99%

mentioning

confidence: 99%

Structural engineering of the HIV‐1 protease molecule with a β‐turn mimic of fixed geometry

1993

View full text Add to dashboard Cite

show abstract

“…2,3 Since that time, solid phase peptide synthesis has been integral in the total chemical synthesis of a number of other enzyme molecules including HIV-1 protease (HIV-1 PR) 4 ; a tethered dimer of HIV-1 PR 5 ; 4-oxalocrotonate tautomerase (4-OT ) 6 ; human type II secretory phospholipase A2 (PLA2) 7 ; barnase 8 ; and the electron transport protein cytochrome b562. 9 Unlike Gutte and Merrifield's original groundbreaking stepwise synthesis of RNase A, 2,3 all of these enzyme syntheses (except for the synthesis of the 62 residue 4-OT 6 ) have relied on some sort of peptide ligation chemistry in addition to solid phase peptide synthesis.…”

Section: Introductionmentioning

confidence: 99%

Total synthesis by modern chemical ligation methods and high resolution (1.1 Å) X‐ray structure of ribonuclease A

2008

View full text Add to dashboard Cite

The total chemical synthesis of RNase A using modern chemical ligation methods is described, illustrating the significant advances that have been made in chemical protein synthesis since Gutte and Merrifield's pioneering preparation of RNase A in 1969. The identity of the synthetic product was confirmed through rigorous characterization, including the determination of the X‐ray crystal structure to 1.1 Angstrom resolution. © 2007 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 90:278–286, 2008.This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com

show abstract

“…Its crystal structure [9,10] confirms those predictions. Other divergences of HIV-l protease are the higher pH for optimal activity f7,11] compared to pepsins, and its much weaker inhibition by pepstatin A [4,7,12] (table 1). Those characteristics are generally shared by renin [13,14]* as positions of oxygens from water molecules [18,19].…”

Section: I~roductionmentioning

confidence: 99%

“…Its crystal structure [9,10] confirms those predictions. Other divergences of HIV-l protease are the higher pH for optimal activity f7,11] compared to pepsins, and its much weaker inhibition by pepstatin A [4,7,12] Fig.1 shows the alignment of 8 active site residues which are crucial for maintaining the hydrogen-bonded structure of the catalytic centers. The role of the fourth residue of each partial sequence (which is mutated in h. renin and retroviral proteases compared to pepsins) in regulating the properties of active site aspartates has rarely been addressed and the source of optimal pH variations among the AP is poorly understood [20].…”

mentioning

confidence: 99%

Modulation of the affinity of aspartic proteases by the mutated residues in active site models

Goldblum

1990

FEBS Letters

View full text Add to dashboard Cite

The active sites of 3 types of aspartic proteases are modeled, based on crystallographic coordinates of endothiapepsin and of a model of HIV-l protease. The enthalpies of deprotonation from neutral to mono-anion and to dianion are calculated with semiempirical minimal neglect of differential overlap, hydrogen bonding corrected (MNDO/H). This quantum mechanical study of models for the active sites of pepsins, human renin and retroviral aspartic proteases demonstrates that the replacements of Thr-218 from pepsins by Ala in human renin and of both Ser-35 and Thr-218 by alanines in retroviral proteases increases the proton affinity and modulates the charge distribution of those active sites compared to the pepsins.Protease, Aspartic; HIV-l; Renin; Pepsin; Model, molecular I~RODUCTIONAPs are a family of endopeptidases which have many common features [l J in addition to structural homology [2]. The catalytic machinery includes two closely positioned aspartic acids with highly conserved neighbor residues. Most APs are maximally active at low pH ('acid proteases'). They have diverse substrate specificities but are inhibited, to varying extents, by a common inhibitor, pepstatin A [3-51. APs are known to be involved in a few disease conditions such as AIDS, essential h~rtension, several cancers and others f6] and are now a major target for designing inhibitors.Recent research establishes that HIV-I protease, which processes the gag and pol polyprotein precursors towards the production of mature viruses, is an AP [7]. Like other retroviral proteases, it has a much shorter polypeptide chain than pepsins and renins, and was postulated to function in dimeric form [B]. Its crystal structure [9,10] confirms those predictions. Other divergences of HIV-l protease are the higher pH for optimal activity f7,11] compared to pepsins, and its much weaker inhibition by pepstatin A [4,7,12] Fig.1 shows the alignment of 8 active site residues which are crucial for maintaining the hydrogen-bonded structure of the catalytic centers. The role of the fourth residue of each partial sequence (which is mutated in h. renin and retroviral proteases compared to pepsins) in regulating the properties of active site aspartates has rarely been addressed and the source of optimal pH variations among the AP is poorly understood [20]. The conservation of spatial relations of those residues in the crystal structures of many native and inhibited enzymes supports the conclusion that it is possible to model the active site of an AP for which only primary structure is known, by using the published coordinates of another. We have taken this approach to study the active sites of h. renin and retroviral proteases based on the published coordinates of endothiapepsin. Models of renin were previously constructed for studying their interactions with inhibitors. However, some details of the structure are missing due to the lack of hydrogen coordinates in X-ray studies of proteins. Some of those proton positions are extremely important for hydrogen bonding.

show abstract

Enzymatic activity of a synthetic 99 residue protein corresponding to the putative HIV-1 protease

Cited by 249 publications

References 25 publications

Structural engineering of the HIV‐1 protease molecule with a β‐turn mimic of fixed geometry

Structural engineering of the HIV‐1 protease molecule with a β‐turn mimic of fixed geometry

Total synthesis by modern chemical ligation methods and high resolution (1.1 Å) X‐ray structure of ribonuclease A

Modulation of the affinity of aspartic proteases by the mutated residues in active site models

Contact Info

Product

Resources

About