Cahyo Budiman, Anna Robreth Robert, Jovi Silvester, Ping-Chin Lee, Kamruddin Ahmed, Thean Chor Leow, Azzyati Mohd Padzil, Meiny Suzery, Kazufumi Takano, Asep Awaludin Prihanto
FK506-binding protein 35 from Plasmodium knowlesi (Pk-FKBP35) is a multidomain peptidyl–prolyl cis –trans isomerase (PPIase) considered a potential target for antimalarial drug development. The protein consists of an N-terminal FK506-binding domain (FKBD) and a tetratricopeptide repeat domain (TPRD) containing a calmodulin-binding motif (CBM). The TPRD domain is known to recognize the conserved MEEVD motif located at the C-terminus of heat shock protein 90 (Hsp90). This dataset describes the biochemical and biophysical characterization of Pk-FKBP35 and its interaction with the Hsp90-derived MEEVD motif. Recombinant full-length FKBP35 (FL-FKBP35) and three variants, namely Pk-FKBD, Pk-TPRD+, and a calmodulin-binding motif deletion mutant (del-CBM), were heterologously expressed in E. coli and purified to high purity. Circular dichroism spectroscopy confirmed the structural integrity of the recombinant proteins. PPIase activity assays using a fluorogenic substrate demonstrated that FL-FKBP35, Pk-FKBD, and del-CBM exhibited catalytic activity, with kcat /KM values of 5.04 ± 0.21 × 10–5, 4.81 ± 0.34 × 10–5, and 5.17 ± 0.17 × 10–5 M-1 s-1, respectively, whereas Pk-TPRD+ showed no detectable activity. Interactions between FKBP35 proteins and the MEEVD peptide were examined using surface plasmon resonance analysis, revealing that the dissociation constant (KD) of FL-FKBP35 (54.75 ± 5.01 µM) was comparable (P > 0.05) to that of Pk-TPRD+ (46.41 ± 5.36 µM), while Pk-FKBD alone showed no detectable binding. Functional assays further demonstrated that MEEVD binding did not significantly affect the intrinsic PPIase activity of FKBP35 (P > 0.05). These data provide detailed biochemical, biophysical, and interaction profiles of Pk-FKBP35 and its interaction with the Hsp90-derived MEEVD motif. © 2026 The Author(s).
Biotechnology Research Institute, University Malaysia Sabah, Jalan UMS, Sabah, Kota Kinabalu, 88400, Malaysia; Faculty Science and Technology, University Malaysia Sabah, Jalan UMS, Sabah, Kota Kinabalu, 88400, Malaysia; Borneo Medical and Health Research Centre, Faculty of Medicine and Health Sciences, Universiti Malaysia Sabah, Jalan UMS, Sabah, Kota Kinabalu, 88400, Malaysia; Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Selangor, UPM Serdang, 43400, Malaysia; Structural Biology and Functional Omics, Malaysia Genome and Vaccine Institute, National Institutes of Biotechnology Malaysia, Selangor, Kajang, 43000, Malaysia; Department of Chemistry, Faculty of Science and Mathematics Universitas Diponegoro, Jl. Prof. Soedarto No.50275, Semarang, 50275, Indonesia; Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, Kyoto, 606-8522, Japan; Faculty of Fisheries and Marine Sciences, Universitas Brawijaya, Jln. Veteran Malang, Jawa Timur, 65145, Indonesia
