In Vivo Dissection of the Intrinsically Disordered Receptor Domain of Tim23

Umut Günsel, Eyal Paz, Ruhita Gupta, Isabella Mathes, Abdussalam Azem, Dejana Mokranjac*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

9 Scopus citations


In the intermembrane space (IMS) of mitochondria, the receptor domain of Tim23 has an essential role during translocation of hundreds of different proteins from the cytosol via the TOM and TIM23 complexes in the outer and inner membranes, respectively. This intrinsically disordered domain, which can even extend into the cytosol, was shown, mostly in vitro, to interact with several subunits of the TOM and TIM23 complexes. To obtain molecular understanding of this organizational hub in the IMS, we dissected the IMS domain of Tim23 in vivo. We show that the interaction surface of Tim23 with Tim50 is larger than previously thought and reveal an unexpected interaction of Tim23 with Pam17 in the IMS, impairment of which influences their interaction in the matrix. Furthermore, mutations of two conserved negatively charged residues of Tim23, close to the inner membrane, prevented dimerization of Tim23. The same mutations increased exposure of Tim23 on the mitochondrial surface, whereas dissipation of membrane potential decreased it. Our results reveal an intricate network of Tim23 interactions in the IMS, whose influence is transduced across two mitochondrial membranes, ensuring efficient translocation of proteins into mitochondria.

Original languageEnglish
Pages (from-to)3326-3337
Number of pages12
JournalJournal of Molecular Biology
Issue number10
StatePublished - 1 May 2020


FundersFunder number
Friedrich-Baur Stiftung
Joint Research Projects on Biophysics LMU-TAU
Petra Robisch and Zdenka Stanic
Scripps Research Institute
Deutsche ForschungsgemeinschaftMO1944/1-1, MO1944/1-2
Israel Science Foundation1389/18


    • TIM23 complex
    • mitochondria
    • presequence pathway
    • protein sorting
    • protein translocation


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