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Selectivity in substrate–enzyme complexation studied by surface forces measurement

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Abstract

The selectivity of substrate in substrate–enzyme complexation of heptaprenyl diphosphate synthase was directly investigated using colloidal probe atomic force microscopy (AFM). This enzyme is composed of two dissociable subunits, which exhibits a catalytic activity only when they are associated together in the presence of a cofactor, Mg2+, and a substrate, farnesyl diphosphate (FPP). We have recently succeeded to directly demonstrate a specific interaction involved in this enzyme reaction and obtain new insights into the molecular mechanism of the reaction using the approach based on the colloidal probe AFM. The AFM measurement showed the adhesive force between the subunits only in the presence of both Mg2+ and FPP. In this study, we studied the substrate selectivity in the complexation by monitoring the adhesive force. The substrates studied are pyrophosphate (PPi), isopentenyl diphosphate (IPP), geranyl diphosphate (GPP), farnesyl monophosphate (FP), and farnesyl geranyl diphosphate (FGPP). No adhesion was observed in the case of PPi, IPP, and GPP. On the other hand, the significant adhesion was observed for phosphate derivatives, which bear prenyl units longer than three. This is in good agreement with the selectivity of the substrates by this enzyme, which catalyzes the condensation reaction of four IPP molecules with FPP to give heptaprenyl (C35) diphosphates. Our study showed a useful methodology for examining the elemental processes of biological reactions.

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Acknowledgment

This work was partially supported by Grant-in-Aids for 21st century COE Research, Giant Molecules and Complex Systems, from the Ministry of Education, Culture, Sports, Science, and Technology of Japan and supported by the CREST program of Japan Science and Technology Agent.

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  1. Takehiro Suzuki

    Present address: National Institute for Environmental Studies, 16–2 Onogawa, Tsukuba-City, Ibaraki, 305-8506, Japan

Authors and Affiliations

  1. Institute for Multidisciplinary Research for Advanced Materials, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai, 980-8577, Japan

    Takehiro Suzuki, Tanetoshi Koyama & Kazue Kurihara

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  1. Takehiro Suzuki
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  2. Tanetoshi Koyama
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  3. Kazue Kurihara
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Correspondence to Kazue Kurihara.

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Suzuki, T., Koyama, T. & Kurihara, K. Selectivity in substrate–enzyme complexation studied by surface forces measurement. Colloid Polym Sci 286, 107–112 (2008). https://doi.org/10.1007/s00396-007-1746-1

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  • DOI: https://doi.org/10.1007/s00396-007-1746-1

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