TY - JOUR
T1 - Structural basis of enzyme activity regulation by the propeptide of L-lysine α-oxidase precursor from Trichoderma viride
AU - Kitagawa, Masaki
AU - Ito, Nanako
AU - Matsumoto, Yuya
AU - Saito, Masaya
AU - Tamura, Takashi
AU - Kusakabe, Hitoshi
AU - Inagaki, Kenji
AU - Imada, Katsumi
N1 - Funding Information:
We thank SPring-8 beamline staffs for technical help in use of beamline BL41XU. This work was supported by JSPS KAKENHI Grant Numbers 24560962 (to K. Inagaki).
Publisher Copyright:
© 2021 The Author(s)
PY - 2021/1
Y1 - 2021/1
N2 - Harmuful proteins are usually synthesized as inactive precursors and are activated by proteolytic processing. L-Amino acid oxidase (LAAO) is a flavoenzyme that catalyzes the oxidative deamination of L-amino acid to produce a 2-oxo acid with ammonia and highly toxic hydrogen peroxide and, therefore, is expressed as a precursor. The LAAO precursor shows significant variation in size and the cleavage pattern for activation. However, the molecular mechanism of how the propeptide suppresses the enzyme activity remains unclear except for deaminating/decarboxylating Pseudomonas L-phenylalanine oxidase (PAO), which has a short N-terminal propeptide composed of 14 residues. Here we show the inactivation mechanism of the L-lysine oxidase (LysOX) precursor (prLysOX), which has a long N-terminal propeptide composed of 77 residues, based on the crystal structure at 1.97 Å resolution. The propeptide of prLysOX indirectly changes the active site structure to inhibit the enzyme activity. prLysOX retains weak enzymatic activity with strict specificity for L-lysine and shows raised activity in acidic conditions. The structures of prLysOX crystals that soaked in a solution with various concentrations of L-lysine have revealed that prLysOX can adopt two conformations; one is the inhibitory form, and the other is very similar to mature LysOX. The propeptide region of the latter form is disordered, and L-lysine is bound to the latter form. These results indicate that prLysOX uses a different strategy from PAO to suppress the enzyme activity and suggest that prLysOX can be activated quickly in response to the environmental change without proteolytic processing.
AB - Harmuful proteins are usually synthesized as inactive precursors and are activated by proteolytic processing. L-Amino acid oxidase (LAAO) is a flavoenzyme that catalyzes the oxidative deamination of L-amino acid to produce a 2-oxo acid with ammonia and highly toxic hydrogen peroxide and, therefore, is expressed as a precursor. The LAAO precursor shows significant variation in size and the cleavage pattern for activation. However, the molecular mechanism of how the propeptide suppresses the enzyme activity remains unclear except for deaminating/decarboxylating Pseudomonas L-phenylalanine oxidase (PAO), which has a short N-terminal propeptide composed of 14 residues. Here we show the inactivation mechanism of the L-lysine oxidase (LysOX) precursor (prLysOX), which has a long N-terminal propeptide composed of 77 residues, based on the crystal structure at 1.97 Å resolution. The propeptide of prLysOX indirectly changes the active site structure to inhibit the enzyme activity. prLysOX retains weak enzymatic activity with strict specificity for L-lysine and shows raised activity in acidic conditions. The structures of prLysOX crystals that soaked in a solution with various concentrations of L-lysine have revealed that prLysOX can adopt two conformations; one is the inhibitory form, and the other is very similar to mature LysOX. The propeptide region of the latter form is disordered, and L-lysine is bound to the latter form. These results indicate that prLysOX uses a different strategy from PAO to suppress the enzyme activity and suggest that prLysOX can be activated quickly in response to the environmental change without proteolytic processing.
KW - Crystal structure
KW - L-Lysine α-oxidase
KW - Precursor
KW - Substrate recognition
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U2 - 10.1016/j.yjsbx.2021.100044
DO - 10.1016/j.yjsbx.2021.100044
M3 - Article
AN - SCOPUS:85099840813
VL - 5
JO - Journal of Structural Biology: X
JF - Journal of Structural Biology: X
SN - 2590-1524
M1 - 100044
ER -