TY - JOUR
T1 - NMR identification of the Tom20 binding segment in mitochondrial presequences
AU - Muto, Takanori
AU - Obita, Takayuki
AU - Abe, Yoshito
AU - Shodai, Toshihiro
AU - Endo, Toshiya
AU - Kohda, Daisuke
N1 - Funding Information:
We are very grateful to Dr M. Mori (Kumamoto University) for providing the cDNA for rat OTC. This project was supported by Grants in aid for Scientific Research from the Ministry of Education, Science and Culture of Japan (T. E.).
PY - 2001/2/16
Y1 - 2001/2/16
N2 - Many mitochondrial proteins are synthesized in the cytosol as precursors with N-terminal presequences, and are imported into mitochondria with the aid of translocator protein complexes containing presequence-binding proteins. Tom20, a receptor protein which functions in an early step of the mitochondrial protein import, recognizes presequences with divergent amino acid sequences. Here, we report the identification of the segments involved in binding to Tom20 in mitochondrial presequences. We monitored the chemical shift perturbation of the NMR signals of five different 15N-labeled presequence peptides by the addition of the cytosolic receptor domain of rat or yeast Tom20. The perturbed segments occupy different positions, either near the N terminus or at the C terminus, in the presequences. Spin label experiments revealed that this is not due to different orientations of the presequence peptides bound to Tom20. The results presented here will offer a starting point to perform detailed analyses of Tom20-binding elements by systematic amino acid replacements.
AB - Many mitochondrial proteins are synthesized in the cytosol as precursors with N-terminal presequences, and are imported into mitochondria with the aid of translocator protein complexes containing presequence-binding proteins. Tom20, a receptor protein which functions in an early step of the mitochondrial protein import, recognizes presequences with divergent amino acid sequences. Here, we report the identification of the segments involved in binding to Tom20 in mitochondrial presequences. We monitored the chemical shift perturbation of the NMR signals of five different 15N-labeled presequence peptides by the addition of the cytosolic receptor domain of rat or yeast Tom20. The perturbed segments occupy different positions, either near the N terminus or at the C terminus, in the presequences. Spin label experiments revealed that this is not due to different orientations of the presequence peptides bound to Tom20. The results presented here will offer a starting point to perform detailed analyses of Tom20-binding elements by systematic amino acid replacements.
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U2 - 10.1006/jmbi.2000.4397
DO - 10.1006/jmbi.2000.4397
M3 - Article
C2 - 11237589
AN - SCOPUS:0035895431
VL - 306
SP - 137
EP - 143
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
SN - 0022-2836
IS - 2
ER -