3LX9

Interconversion of Human Lysosomal Enzyme Specificities

Summary for 3LX9

• Entry DOI: 10.2210/pdb3lx9/pdb
• Related: 1R46 3HG2 3LXA 3LXB 3LXC
• Descriptor: Alpha-galactosidase A, alpha-D-mannopyranose-(1-3)-beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose, alpha-D-mannopyranose-(1-3)-beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-[alpha-L-fucopyranose-(1-3)]2-acetamido-2-deoxy-beta-D-glucopyranose, ... (7 entities in total)
• Functional Keywords: glycoprotein, carbohydrate-binding protein, glycosidase, lysosomal enzyme, (beta/alpha)8 barrel, enzyme interconversion, disease mutation, disulfide bond, hydrolase, lysosome
• Biological source: Homo sapiens (human)
• Total number of polymer chains: 2
• Total formula weight: 96142.16

Authors

Tomasic, I.B.,Metcalf, M.C.,Guce, A.I.,Clark, N.E.,Garman, S.C. (deposition date: 2010-02-25, release date: 2010-05-05, Last modification date: 2024-11-20)

Primary citation

Tomasic, I.B.,Metcalf, M.C.,Guce, A.I.,Clark, N.E.,Garman, S.C.
Interconversion of the specificities of human lysosomal enzymes associated with Fabry and Schindler diseases.
J.Biol.Chem., 285:21560-21566, 2010

PubMed Abstract: The human lysosomal enzymes alpha-galactosidase (alpha-GAL, EC 3.2.1.22) and alpha-N-acetylgalactosaminidase (alpha-NAGAL, EC 3.2.1.49) share 46% amino acid sequence identity and have similar folds. The active sites of the two enzymes share 11 of 13 amino acids, differing only where they interact with the 2-position of the substrates. Using a rational protein engineering approach, we interconverted the enzymatic specificity of alpha- GAL and alpha-NAGAL. The engineered alpha-GAL (which we call alpha-GAL(SA)) retains the antigenicity of alpha-GAL but has acquired the enzymatic specificity of alpha-NAGAL. Conversely, the engineered alpha-NAGAL (which we call alpha-NAGAL(EL)) retains the antigenicity of alpha-NAGAL but has acquired the enzymatic specificity of the alpha-GAL enzyme. Comparison of the crystal structures of the designed enzyme alpha-GAL(SA) to the wild-type enzymes shows that active sites of alpha-GAL(SA) and alpha-NAGAL superimpose well, indicating success of the rational design. The designed enzymes might be useful as non-immunogenic alternatives in enzyme replacement therapy for treatment of lysosomal storage disorders such as Fabry disease.

PubMed: 20444686
DOI: 10.1074/jbc.M110.118588

Experimental method

X-RAY DIFFRACTION (2.04 Å)