Inhibitors of mitochondrial fission and methods of use thereof
The present disclosure provides peptides and constructs that inhibit mitochondrial fission, and compositions comprising the peptides or constructs. The present disclosure provides methods of reducing abnormal mitochondrial fission in a cell. Also provided are methods for designing and validating mitochondrial fission inhibitor constructs and peptides, including but not limited to, evaluating the effects of the constructs and peptides on dynamin- 1-related protein (Drp1) GTPase activity, binding of Drp1 to mitochondrial fission 1 protein (Fis1), reduction of mitochondrial damage, reduction in cell death, inhibition of mitochondrial fragmentation in a cell under pathological conditions, and reduced loss of neurites in primary dopaminergic neurons in a Parkinsonism cell culture.
1. A mitochondrial fission inhibitor peptide construct comprising a mitochondrial fission inhibitor peptide consisting of 7, 8 or 9 amino acids,
wherein the inhibitor peptide comprises the amino acid sequence as set forth in SEQ ID NO:12 having none, one, two or three conservative amino acid substitutions; and
a carrier peptide selected from the group consisting of SEQ ID NOs: 31-45,
wherein the mitochondrial fission inhibitor peptide construct is no more than 62 amino acids in length.
2. The inhibitor peptide construct of claim 1 , wherein the one, two or three amino acid substitutions are selected from the following:
the aspartic acid at position 1 of SEQ ID NO:12 is substituted by glutamic acid,
the leucine at position 2 of SEQ ID NO:12 is substituted by isoleucine, methionine, valine, or phenylalanine,
the leucine at position 3 of SEQ ID NO:12 is substituted by isoleucine, methionine, or phenylalanine,
the arginine at position 5 of SEQ ID NO:12 is substituted by lysine,
the glycine at position 6 of SEQ ID NO:12 is substituted by alanine, threonine or serine, and
the serine at position 7 of SEQ ID NO:12 is substituted by glycine, alanine or threonine.
3. The inhibitor peptide construct of claim 1 , further comprising a linker, wherein the linker is positioned between the fission inhibitor peptide and the carrier peptide, and wherein the linker is selected from the group consisting of G, GG, GGG and GGGG (SEQ ID NO:62).
4. The mitochondrial fission inhibitor peptide construct of claim 1 , wherein the fission inhibitor peptide consists of 8 amino acids.
5. The inhibitor peptide construct of claim 4 , wherein the carrier peptide is SEQ ID NO:32 or SEQ ID NO:33.
6. The inhibitor peptide construct of claim 5 , further comprising a linker, wherein the linker is positioned between the fission inhibitor peptide and the carrier peptide, and wherein the linker is selected from the group consisting of G, GG, GGG and GGGG (SEQ ID NO:62).
7. The inhibitor peptide construct of claim 4 , wherein the one, two or three amino acid substitutions are selected from the following:
the aspartic acid at position 1 of SEQ ID NO:12 is substituted by glutamic acid,
the leucine at position 2 of SEQ ID NO:12 is substituted by isoleucine, methionine, valine, or phenylalanine,
the leucine at position 3 of SEQ ID NO:12 is substituted by isoleucine, methionine, or phenylalanine,
the arginine at position 5 of SEQ ID NO:12 is substituted by lysine,
the glycine at position 6 of SEQ ID NO:12 is substituted by alanine, threonine or serine, and
the serine at position 7 of SEQ ID NO:12 is substituted by glycine, alanine or threonine.
8. The inhibitor peptide construct of claim 4 , wherein the fission inhibitor peptide comprises SEQ ID NO:12.
9. The inhibitor peptide construct of claim 1 , wherein the inhibitor peptide construct is linear.
10. The inhibitor construct of claim 1 , wherein the inhibitor peptide construct is cyclic.