Crystal structure of human alpha-N-acetylglucosaminidase
View Patent ↗The present invention provides the three-dimensional structure of human α-N-acetylglucosaminidase (NAGLU) protein. This crystallographic information is useful in the identification and development of novel binding compounds of NAGLU, NAGLU mutants, for example, those associated with Sanfilippo syndrome type B (mucopolysaccharidosis III B (MPS III-B)), and other NAGLU family members (family 89 α-N-acetylglucosaminidase) which may modulate the activity and/or stability of mutated NAGLU. Such compounds may be useful for the treatment of Sanfilippo syndrome type B (mucopolysaccharidosis III B (MPS III-B)).
1. A method of selecting or designing a α-N-acetylglucosaminidase (NAGLU) polypeptide binding compound, the method comprising:
a) displaying the atomic coordinates of amino acids 134-658 as set forth in Table 3.1, Table 3.2, or Table 3.3 to form a three-dimensional structure of a first NAGLU polypeptide using a programmed computer;
b) constructing a model of a second NAGLU polypeptide using the three-dimensional structure of the first NAGLU polypeptide as a template, wherein the second NAGLU polypeptide differs from the first NAGLU polypeptide in that the second NAGLU polypeptide has at least one amino acid substitution, deletion or duplication not present in the first NAGLU polypeptide;
c) screening a binding compound that binds to the second NAGLU polypeptide in vitro, or in vivo for particular characteristics or desired properties; and
d) selecting or designing the binding compound with particular characteristics or desired properties.
2. The method of claim 1 , wherein the position of the amino acid substitution, deletion or duplication of the second NAGLU polypeptide is first identified in the three-dimensional structure of the first NAGLU polypeptide, and wherein the amino acid substitution, deletion or duplication in the second NAGLU polypeptide is selected based on a known amino acid substitution, deletion or duplication that is associated with or leads to mucopolysaccharidosis III B (MPS III-B).
3. The method of claim 2 , wherein the amino acid substitution, deletion, or duplication that is associated with or leads to mucopolysaccharidosis III B (MPS III-B) is selected from the group consisting of the amino acid substitution, deletion, or duplication as set forth in Table 4.
4. A method of selecting or designing a α-N-acetylglucosaminidase (NAGLU) polypeptide binding compound, the method comprising:
a) providing the atomic coordinates of a NAGLU polypeptide as set forth in Table 3.1, Table 3.2, or Table 3.3;
b) displaying the atomic coordinates set forth in Table 3 to form a three-dimensional structure of a first NAGLU polypeptide;
c) constructing a model of a second NAGLU polypeptide using the three-dimensional structure of the first NAGLU polypeptide as a template, wherein the second NAGLU polypeptide differs from the first NAGLU polypeptide in that the second NAGLU polypeptide has at least one amino acid substitution, deletion or duplication not present in the first NAGLU polypeptide; and
d) screening a binding compound that binds to the second NAGLU polypeptide in vitro, or in vivo for particular characteristics or desired properties; and
e) selecting or designing the binding compound with particular characteristics or desired properties.
5. A method of identifying a drug candidate test compound for the treatment of mucopolysaccharidosis III B (MPS III-B), the method comprising:
a) displaying the atomic coordinates as set forth in Table 3.1, Table 3.2, or Table 3.3 to form a three-dimensional structure of a first NAGLU polypeptide using a programmed computer;
b) constructing a model of a second NAGLU polypeptide using the three-dimensional structure of the first NAGLU polypeptide as a template, wherein the second NAGLU polypeptide differs from the first NAGLU polypeptide in that the second NAGLU polypeptide has at least one amino acid substitution, deletion or duplication not present in the first NAGLU polypeptide, and wherein the position of the amino acid substitution, deletion or duplication of the second NAGLU polypeptide is first identified in three-dimensional structure of the first NAGLU polypeptide, and wherein the amino acid substitution, deletion or duplication in the second NAGLU polypeptide is selected based on a known amino acid substitution, deletion or duplication that is associated with or leads to mucopolysaccharidosis III B (MPS III-B);
c) selecting a test compound having the best fit with the NAGLU polypeptide comprising one or more amino acid substitution, deletion or duplication; and
d) assaying in vivo or in vitro the ability of the test compound to modulate NAGLU enzyme activity, modulate stability and/or modulate intracellular trafficking of the NAGLU polypeptide comprising one or more amino acid mutations, wherein a test compound that modulates NAGLU enzyme activity, stability and/or intracellular trafficking of the NAGLU polypeptide comprising one or more amino acid mutations is considered a drug candidate compound for treating MPS III-B.