| Full text | |
| Author(s): |
Damasceno, Ticiane F.
[1]
;
Dias, Renata O.
[1]
;
de Oliveira, Juliana R.
[2]
;
Salinas, Roberto K.
[1]
;
Juliano, Maria A.
[2]
;
Ferreira, Clelia
[1]
;
Terra, Walter R.
[1]
Total Authors: 7
|
| Affiliation: | [1] Univ Sao Paulo, Inst Quim, Dept Bioquim, Ave Prof Lineu Prestes 748, BR-05508000 Sao Paulo - Brazil
[2] Univ Fed Sao Paulo, Dept Biofis, Rua Tres de Maio 100, BR-04044020 Sao Paulo - Brazil
Total Affiliations: 2
|
| Document type: | Journal article |
| Source: | Insect Biochemistry and Molecular Biology; v. 89, p. 17-30, OCT 2017. |
| Web of Science Citations: | 3 |
| Abstract | |
Cathepsins L are the major digestive peptidases in the beetle Tenebrio molitor. Two digestive cathepsins L (TmCAL2 and TmCAL3) from it had their 3D structures solved. The aim of this paper was to study in details TmCAL3 specificity and properties and relate them to its 3D structure. Recombinant TmCAL3 was assayed with 64 oligopeptides with different amino acid replacements in positions P2, P1, P1' and P2'. Results showed that TmCAL3 S2 specificity differs from the human enzyme and that its specificities also explain why on autoactivation two propeptide residues remain in the enzyme. Data on free energy of binding and of activation showed that S1 and S2' are mainly involved in substrate binding, S1' acts in substrate binding and catalysis, whereas 52 is implied mainly in catalysis. Enzyme subsite residues were identified by docking with the same oligopeptide used for kinetics. The subsite hydrophobicities were calculated from the efficiency of hydrolysis of different amino acid replacements in the peptide and from docking data. The results were closer for S1 and S2' than for SI and S2, indicating that the residue subsites that were more involved in transition state binding are different from those binding the substrate seen in docking. Besides TmCAL1-3, there are nine other cathepsins L, most of them more expressed at midgut. They are supposed to be directed to lysosomes by a Drosophila-like Lerp receptor and/or motifs in their prodomains. The mannose 6-phosphate lysosomal sorting machinery is absent from T. molitor transcriptome. Cathepsin L direction to midgut contents seems to depend on overexpression. 2017 Elsevier Ltd. All rights reserved. (AU) | |
| FAPESP's process: | 15/19541-7 - Evolution of molecular processes associated with nutrient absorption, fluid circulation and buffering in the intestines of Hemiptera and Diptera |
| Grantee: | Renata de Oliveira Dias |
| Support Opportunities: | Scholarships in Brazil - Post-Doctoral |
| FAPESP's process: | 16/07490-1 - Structural studies of the Na+/Ca2+ exchanger and of proteins from the Zika virus by Nuclear Magnetic Resonance spectroscopy in solution |
| Grantee: | Roberto Kopke Salinas |
| Support Opportunities: | Regular Research Grants |
| FAPESP's process: | 11/51685-8 - Insect digestion: a molecular, cellular, physiological and evolutionary approach |
| Grantee: | Walter Ribeiro Terra |
| Support Opportunities: | Research Projects - Thematic Grants |