Assay Systems for HIV-1 Proteinase and Their Use for Evaluation of Inhibitors

Billich, Andreas; Billich, S.; Rosenwirth, Brigitte

doi:10.1177/095632029100200201

Cited by 8 publications

(5 citation statements)

References 72 publications

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“…Table I summarizes the effect of different types of salt on the kinetics of HIV-1 PR. In accordance with studies using NaCI [10,11] and ammonium sulfate [10], we found that the enzyme is more active at a high ionic ~CH=COO, Na=SO4 and NaCI were assayed at a total ionic strenitth of 2,57 M in buffer conlain. ins 0,25 M potassium phosphate, pH 6.$.…”

Section: Resultssupporting

confidence: 86%

“…Recombinant PR or chemically synthesized PR have been purified and used as sources for investigation of enzymatic properties and inhibitor studies (for review see [5][6][7][8][9]). High ionic strength was found to optimize the enzymatic activity by lowering the Michaelis Menten constant (K,0; the turnover number (Kent) was not affected [10,11].…”

Section: Introductionmentioning

confidence: 99%

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The effect of salt on the Michaelis Menten constant of the HIV‐1 protease correlates with the Hofmeister series

1991

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The effect of different typ~s of salt on the proteolytic activity of H IV. I protease was studied, At a similar ionic strength, the enzyme activity chanlled according to the saltintl out effect of the =ons used (I.lofmeister ~rles), Kinetic studies showed that a stronger saltln$ out effect of tit© ions rather than the higher ionic strength I~r sc increased the affinity to the substrate (K..) but in general did not alter the K.., value, H IV. I pretense; Enzyme kinetics: Salt effect: Hofmeister series

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Section: Resultssupporting

confidence: 86%

Section: Introductionmentioning

confidence: 99%

The effect of salt on the Michaelis Menten constant of the HIV‐1 protease correlates with the Hofmeister series

1991

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“…Cellular fatty acid-binding proteins are a highly conserved family of proteins involved in intracellular fatty acid metabolism and trafficking. It has been suggested that in brain and heart, B-FABP and H-FABP regulate the supply of fatty acids to the mitochondria for ß-oxidation (29,30). The mammary gland, however, is a highly lipogenic tissue, and fatty acids are not likely to be a major fuel for its metabolism.…”

Section: Discussionmentioning

confidence: 99%

Breast Cancer Prevention by Hormonally Induced Mammary Gland Differentiation: The Role of a Novel Mammary Growth Inhibitor and Differentiation Factor MRG

Shi¹

2001

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Public reporting burden for this collection of information is estimated to average 1 hour per response, including the time for reviewing instructions, searching existing data sources, gathering and maintaining the data needed and completing and reviewing this collection of information. Send comments regarding this burden estimate or any other aspect of this collection of information, including suggestions for reducing this burden to Washington Headquarters Services, Directorate for Information Operations and Reports, 1215 Jefferson Davis Highway, Suite 1204. Arlington, VA 22202-4302, and PERFORMING ORGANIZATION NAME(S) AND ADDRESS(ES)Long Island Jewish Medical Center New Hyde Park, New York 11040 E-Mail: shi@lij.edu SPONSORING / MONITORING AGENCY NAME(S) AND ADDRESS(ES)U AbstractWe have previously identified and characterized a novel tumor growth inhibitor and a fatty acid binding protein in human mammary gland and named it as Mammary derived growth inhibitor Related Gene MRG. MRG has tumor-suppressing activities; it inhibits breast cancer cell growth in vitro and tumor growth in vivo. Here, the effects of MRG on mammary gland differentiation and its interaction with o>3 polyunsaturated fatty acids (o>3 PUFA) on growth inhibition were investigated. MRG protein expression was associated with human mammary gland differentiation with the highest expression observed in the differentiated alveolar mammary epithelial cells from the lactating gland. Overexpression of MRG in human breast cancer cells induced differentiation with changes in cellular morphology and a significant increase in the production of lipid droplets. Treatment of mouse mammary gland in organ culture with MRG protein resulted in a differentiated morphology and stimulation of ß-casein expression. Treatment of human breast cancer cells with u>3 PUFA DHA resulted in a differential growth inhibition proportional to their MRG expression. MRG transfected cells or MRG protein treated cells were much more sensitive to DHA-induced growth inhibition compared with MRG negative or control non-treated cells. Our results suggest that MRG is a candidate mediator of the differentiating effect of pregnancy on breast epithelial cells and may play a major role in u>3 PUFA-mediated tumor suppression.

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“…In general, the lower the Ki, the greater will be the discrepancy. In addition, Ki values for HIV protease inhibitors are known to be lower at higher ionic strengths (3).…”

mentioning

confidence: 99%

“…These active agents contain a nonhydrolyzable, dipeptide isostere such as hydroxyethylene (20,22,41) or hydroxyethylamine (34,37) as an active moiety which mimics the putative transition state of the aspartic protease-catalyzed reaction. Twofold (C2) symmetric inhibitors of HIV protease represent another class of potent HIV protease inhibitors which were created by Erickson et al on the basis of the three-dimensional symmetry of the enzyme active site (5 3'-azido-2',3'-dideoxythymidine [AZT], 2',3'-dideoxyinosine [ddI], and 2',3'-dideoxycytidine) will likely enhance the efficacy and/or reduce the toxic effects of each drug. Indeed, we have observed synergism against HIV-1 between certain C2 symmetric HIV inhibitors and AZT (13).…”

mentioning

confidence: 99%

In vitro anti-human immunodeficiency virus (HIV) activities of transition state mimetic HIV protease inhibitors containing allophenylnorstatine

Kageyama

Mimoto

Murakawa

et al. 1993

Antimicrob Agents Chemother

117

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Transition state mimetic tripeptide human immunodeficiency virus (HIV) protease inhibitors containing allophenylnorstatine [(2S,3S)-3-amino-2-hydroxy-4-phenylbutyric acid] were synthesized and tested for activity against HIV in vitro. Two compounds, KNI-227 and KNI-272, which were highly potent against HIV protease with little inhibition of other aspartic proteases, showed the most potent activity against the infectivity and cytopathic effect of a wide spectrum of HIV strains. As tested in target CD4+ ATH8 cells, the 50% inhibitory concentrations of KNI-227 against HIV type 1 LAI (HIV-1LAI), HIV-1RF, HIV-1MN, and HIV-2ROD were 0.1, 0.02, 0.03, and 0.1 microM, respectively, while those of KNI-272 were 0.1, 0.02, 0.04, and 0.1 microM, respectively. Both agents completely blocked the replication of 3'-azido-2',3'-dideoxythymidine-sensitive and -insensitive clinical HIV-1 isolates at 0.08 microM as tested in target phytohemagglutinin-activated peripheral blood mononuclear cells. The ratios of 50% cytotoxic concentrations to 50% inhibitory concentrations for KNI-227 and KNI-272 were approximately 2,500 and > 4,000, respectively, as assessed in peripheral blood mononuclear cells. Both compounds blocked the posttranslational cleavage of the p55 precursor protein to generate the mature p24 Gag protein in stably HIV-1-infected cells. The n-octanol-water partition coefficients of KNI-227 and KNI-272 were high, with log Po/w values of 3.79 and 3.56, respectively. Degradation of KNI-227 and KNI-272 in the presence of pepsin (1 mg/ml, pH 2.2) at 37 degrees C for 24 h was negligible. Current data warrant further careful investigations toward possible clinical application of these two novel compounds.

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Assay Systems for HIV-1 Proteinase and Their Use for Evaluation of Inhibitors

Cited by 8 publications

References 72 publications

The effect of salt on the Michaelis Menten constant of the HIV‐1 protease correlates with the Hofmeister series

The effect of salt on the Michaelis Menten constant of the HIV‐1 protease correlates with the Hofmeister series

Breast Cancer Prevention by Hormonally Induced Mammary Gland Differentiation: The Role of a Novel Mammary Growth Inhibitor and Differentiation Factor MRG

In vitro anti-human immunodeficiency virus (HIV) activities of transition state mimetic HIV protease inhibitors containing allophenylnorstatine

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