Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.2.1.23 (beta-galactosidase)
 14,648 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The "protective protein" is a glycoprotein that associates with lysosomal beta-galactosidase and neuraminidase and is deficient in the autosomal recessive disorder galactosialidosis. We have isolated the cDNA encoding human "protective protein". The clone recognizes a 2 kb mRNA in normal cells that is not evident in fibroblasts of an early infantile galactosialidosis patient. The cDNA directs the synthesis of a 452 amino acid precursor molecule that is processed in vivo to yield mature "protective protein," a heterodimer of 32 kd and 20 kd polypeptides held together by disulfide bridges. This mature form is also biologically functional since it restores beta-galactosidase and neuraminidase activities in galactosialidosis cells. The predicted amino acid sequence of the "protective protein" bears homology to yeast carboxypeptidase Y and the KEX1 gene product. This suggests a protease activity for the "protective protein."
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PMID:Expression of cDNA encoding the human "protective protein" associated with lysosomal beta-galactosidase and neuraminidase: homology to yeast proteases. 313 30

The deficiency of the lysosomal protective protein/carboxypeptidase L (CARB L) causes the lysosomal storage disorder, galactosialidosis, characterized by neuraminidase and beta-galactosidase deficiencies in patients' cells. The three enzymes form a complex inside the lysosome, and the neuraminidase and beta-galactosidase deficiencies are secondary to CARB L deficiency. Sequence similarity and common enzymological properties suggest that the protomeric tertiary structure of CARB L is conserved within a family of serine carboxypeptidases which includes the yeast carboxypeptidase Y, killer expression I gene product and several plant carboxypeptidases. We used this homology to build a model of the CARB L structure based on the recently published X-ray atomic coordinates of the wheat carboxypeptidase II (CPDW-II) which shares 32% primary structure identity with CARB L. Small insertions and deletions were accommodated into the model structure by energy minimization using the DREIDING II force field. The C alpha atomic coordinates of the final CARB L model have a RMS shift of 1.01 A compared to the corresponding conserved residues in the CPDW-II template structure. The correct orientation of the homologous catalytic triad residues Ser150, His429 and Asp392, the potential energy calculations and the distribution of hydrophobic and hydrophillic residues in the structure all support the validity of the CARB L model. Most missense mutations identified in galactosialidosis patients were located in secondary structural elements except for the Tyr211-->Asn mutation which is in a loop. The other mutant residues have their side chains deeply buried in the central beta-sheet of the model structure except for the Phe412-->Val mutation which is located in the dimer interface. The predicted effects of specific mutations on CARB L structural stability correlates well with recently published transient expression studies of mutant CARB L (Shimmoto, M. et al., J. Clin. Invest., 91:2393-2399, 1993).
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PMID:Homologous modeling of the lysosomal protective protein/carboxypeptidase L: structural and functional implications of mutations identified in galactosialidosis patients. 814 24

Human cathepsin A ("lysosomal protective protein"; E.C.3.4.16.5) is a multifunctional lysosomal protein which forms a high-molecular-weight complex with beta-galactosidase and alpha-neuraminidase, protecting them against intralysosomal proteolysis. In addition to this protective function, cathepsin A is a serine carboxypeptidase and the understanding of its catalytic function requires a definition of its substrate specificity. For this purpose, we used a combined experimental [Pshezhetsky, A. V., Vinogradova, M. V., Elsliger, M.-A., El-Zein, F., Svedas, V.K., & Potier, M. (1995) Anal. Biochem. 230, 303-307] and theoretical approach comparing cathepsin A to two different homologous carboxypeptidases of the same family: yeast carboxypeptidase Y and wheat carboxypeptidase II. We computed the energies involved in substrate binding to the S1' subsite (C-terminal) of cathepsin A using a structural model based on the X-ray structure of the homologous wheat carboxypeptidase II. The binding energies of N-blocked Phe-Xaa dipeptide substrates to the active sites of cathepsin A, wheat carboxypeptidase II, and yeast carboxypeptidase Y were estimated using a molecular mechanics force field supplemented with a solvation energy term. This theoretical analysis showed a good correlation with the experimentally determined free energies of substrate binding. This result validates the use of this approach to analyze the energetics of substrate binding to the S1' subsite and provides a rational interpretation of serine carboxypeptidase-substrate interactions in molecular terms. We conclude that the three serine carboxypeptidases have similar affinities for substrates with hydrophobic P1' amino acid residues but that the wheat enzyme has an additional capacity for binding positively charged P1' residues. Finally, the substrate specificity of human cathepsin A is very similar to that of carboxypeptidase Y, with a high binding affinity for substrates with hydrophobic P1' residues, but the affinity of cathepsin A for P1; Phe residue is higher than for the Leu residue.
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PMID:Comparative modeling of substrate binding in the S1' subsite of serine carboxypeptidases from yeast, wheat, and human. 894 54

Cathepsin A is a mammalian lysosomal enzyme that catalyzes the hydrolysis of the carboxy-terminal amino acids of polypeptides and also regulates beta-galactosidase and neuraminidase-1 activities through the formation of a multienzymic complex in lysosomes. Human cathepsin A (hCathA), yeast carboxypeptidase (CPY), and wheat carboxypeptidase II (CPW) belong to the alpha/beta-hydrolase fold family. They have structurally similar active-site clefts, but there are small differences in the amino acid residues comprising their active sites that might determine the substrate specificity and sensitivity to microbial inhibitors including chymostatin. To examine the selectivity and binding mechanism of chymostatin as to hCathA, CPY, and CPW at the atomic level, we analyzed the interaction energy between chymostatin and each protein quantitatively by semiempirical molecular orbital calculation AM1 with the continuum solvent model. We predicted the electrostatic repulsion between the P3 cyclic arginine residue of the inhibitor and the Arg344 in the S3 active subsite of hCathA. Genetic conversion of Arg344 of the wild-type hCathA to Ile also caused an increase in its sensitivity to chymostatin, which was correlated with the decrease in the interaction energy calculated with the molecular orbital method. The present results suggest that such molecular calculation should be useful for evaluating the interactions between ligands, including inhibitors and homologous enzymes, in their docking models.
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PMID:Comparative analysis of binding energy of chymostatin with human cathepsin A and its homologous proteins by molecular orbital calculation. 1699 40