Detergent tween 80 modifies the specific activity of PAI-1

Ehnebom, J.; Björquist, Petter; Andersson, Jan-Olof; Johansson, Tomas; Deinum, Johanna

doi:10.1016/s0268-9499(97)80061-6

Cited by 15 publications

(9 citation statements)

References 19 publications

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“…1A, lanes 1-3). We could not observe a "brand"-specific effect of TX-100 (Sigma vs Boehringer) as reported for Tween 80 (Sigma vs Pierce) by Ehnebom et al (42).…”

Section: Discussionsupporting

confidence: 52%

Modulation of Plasminogen Activator Inhibitor 1 by Triton X-100 – Identification of Two Consecutive Conformational Transitions

Gils¹,

Declerck²

1998

Thromb Haemost

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SummaryPlasminogen activator inhibitor-1 (PAI-1) is a unique member of the serpin superfamily because of its conformational and functional flexibility. In the present study, we have evaluated the influence of the nonionic detergent Triton X-100 (TX-100) on the functional stability and conformational transitions of PAI-1.At 37° C, TX-100 induced a concentration-dependent decrease of the functional half-life of PAI-1 resulting in half-lives of 177 ± 54 min (mean ± SD, n = 3), 19 ± 2 min, 1.7 ± 0.3 min and 0.53 ± 0.03 min in the presence of 0.005, 0.010, 0.020 and 0.2% TX-100, respectively, compared to a half-life of 270 ± 146 min in the absence of TX-100. Conformational analysis at various time points and at different temperatures (0° C, 25° C, 37° C) revealed that this inactivation proceeds through the formation of a substrate-like intermediate followed by the formation of the latent form. Kinetic evaluation demonstrated that this conversion fits to two consecutive first-order transitions, i.e. active substrate latent. The k1 value was strongly dependent on the concentration of TX-100 (e.g. 0.002 ± 0.0006 s -1 and 0.029 ± 0.003 s -1 for 0.01% and 0.2% TX-100 at 37° C) whereas the conversion of substrate to latent (k2) was virtually independent of the TX-100 concentration (i.e. 0.012 ± 0.002 s -1 and 0.011 ± 0.001 s -1 for 0.01 and 0.2% TX-100 at 37° C).Experiments with a variety of other non-ionic amphiphilic compounds revealed that the amphiphilic character of the compound is, at least in part, responsible for the observed effects and strongly indicate that the currently reported mechanism of inactivation is of general importance for the conformational transitions in PAI-1.In conclusion, TX-100 changes the initial conformation of PAI-1 resulting in altered functional properties. This observation allows us to develop a new model for the mechanism involved in the conformational flexibility of PAI-1 and may provide new insights for the development of strategies for interference with PAI-1 activity.

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“…1A, lanes 1-3). We could not observe a "brand"-specific effect of TX-100 (Sigma vs Boehringer) as reported for Tween 80 (Sigma vs Pierce) by Ehnebom et al (42).…”

Section: Discussionsupporting

confidence: 52%

Modulation of Plasminogen Activator Inhibitor 1 by Triton X-100 – Identification of Two Consecutive Conformational Transitions

Gils¹,

Declerck²

1998

Thromb Haemost

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“…The running buffer in the direct binding experiments, here referred to as PBSTD buffer, was made by dilution of 10ϫ (BR-1006-72 from Biacore) to 50 mM sodium phosphate, 100 mM NaCl (pH 7.4), and supplemented with 0.005% Tween 20 and 1% DMSO. The anionic detergent Tween 20 has previously been found not to inhibit PAI-1 (14) and was therefore used in all assays to prevent adhesion. Before use in Biacore, the running buffer was degassed for 5 min in an ultrasound bath (Branson 2510).…”

Section: Methodsmentioning

confidence: 99%

Characterization of a Small Molecule Inhibitor of Plasminogen Activator Inhibitor Type 1 That Accelerates the Transition into the Latent Conformation

Fjellström

Deinum

Sjögren

et al. 2013

Journal of Biological Chemistry

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Background: Inhibition of PAI-1 may yield beneficial effects in e.g. cardiovascular diseases and cancer. Results: The small molecule PAI-1 inhibitor, AZ3976, binds latent but not active PAI-1. The structure of the AZ3976⅐latent PAI-1 complex is presented. Conclusion: AZ3976 inhibits PAI-1 by accelerating latency transition, presumably by binding a prelatent form of PAI-1. Significance: This study provides new drug design opportunities for PAI-1 inhibitors.

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“…Partial reactivation of the latent form can be achieved by denaturing agents and subsequent refolding [ 105 ], and also in vivo reactivation of latent PAI-1 has been observed [ 106 ]. The conversion of PAI-1 from the active to the latent conformation appears to be unique among serpins in that it occurs spontaneously at a relatively rapid rate [ 107 , 108 ]. It is believed that latency transition represents a regulatory mechanism that reduces the possibility of thrombosis from a prolonged antifibrinolytic action of PAI-1 [ 14 ].…”

Section: Functional Stability Of Pai-1mentioning

confidence: 99%

Functional Stability of Plasminogen Activator Inhibitor-1

Yıldız

Kuru

Öner

et al. 2014

The Scientific World Journal

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Plasminogen activator inhibitor-1 (PAI-1) is the main inhibitor of plasminogen activators, such as tissue-type plasminogen activator (t-PA) and urokinase-type plasminogen activator (u-PA), and a major regulator of the fibrinolytic system. PAI-1 plays a pivotal role in acute thrombotic events such as deep vein thrombosis (DVT) and myocardial infarction (MI). The biological effects of PAI-1 extend far beyond thrombosis including its critical role in fibrotic disorders, atherosclerosis, renal and pulmonary fibrosis, type-2 diabetes, and cancer. The conversion of PAI-1 from the active to the latent conformation appears to be unique among serpins in that it occurs spontaneously at a relatively rapid rate. Latency transition is believed to represent a regulatory mechanism, reducing the risk of thrombosis from a prolonged antifibrinolytic action of PAI-1. Thus, relying solely on plasma concentrations of PAI-1 without assessing its function may be misleading in interpreting the role of PAI-1 in many complex diseases. Environmental conditions, interaction with other proteins, mutations, and glycosylation are the main factors that have a significant impact on the stability of the PAI-1 structure. This review provides an overview on the current knowledge on PAI-1 especially importance of PAI-1 level and stability and highlights the potential use of PAI-1 inhibitors for treating cardiovascular disease.

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Detergent tween 80 modifies the specific activity of PAI-1

Cited by 15 publications

References 19 publications

Modulation of Plasminogen Activator Inhibitor 1 by Triton X-100 – Identification of Two Consecutive Conformational Transitions

Modulation of Plasminogen Activator Inhibitor 1 by Triton X-100 – Identification of Two Consecutive Conformational Transitions

Characterization of a Small Molecule Inhibitor of Plasminogen Activator Inhibitor Type 1 That Accelerates the Transition into the Latent Conformation

Functional Stability of Plasminogen Activator Inhibitor-1

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