Lupin γ-conglutin and soybean BG7S are two legume seed proteins strongly similar to plant endo-β-glucanases inhibitors acting against fungal GH11 and GH12 glycoside hydrolase. However these proteins lack inhibitory activity. Here we describe the conversion of lupin γ-conglutin to an active inhibitor of endo-β-glucanases belonging to GH11 family. A set of γ-conglutin mutants was designed and expressed in Pichia pastoris, along with the wild-type protein. Unexpectedly, this latter was able to inhibit a GH11 enzyme, but not GH12, whereas the mutants were able to modulate the inhibition capacity. In lupin, γ-conglutin is naturally cleaved in two subunits, whereas in P. pastoris it is not. The lack of proteolytic cleavage is one of the reasons at the basis of the inhibitory activity of recombinant γ-conglutin. The results provide new insights about structural features at the basis of the lack of inhibitory activity of wild-type γ-conglutin and its legume homologues.

Structural basis of the lack of endo-glucanase inhibitory activity of Lupinus albus γ-conglutin / A. Scarafoni, A. Consonni, S. Pessina, S. Balzaretti, J. Capraro, E. Galanti, M. Duranti. - In: PLANT PHYSIOLOGY AND BIOCHEMISTRY. - ISSN 0981-9428. - 99(2016 Feb), pp. 79-85. [10.1016/j.plaphy.2015.11.008]

Structural basis of the lack of endo-glucanase inhibitory activity of Lupinus albus γ-conglutin

A. Scarafoni
Primo
;
A. Consonni
Secondo
;
S. Balzaretti;J. Capraro;E. Galanti
Penultimo
;
M. Duranti
Ultimo
2016

Abstract

Lupin γ-conglutin and soybean BG7S are two legume seed proteins strongly similar to plant endo-β-glucanases inhibitors acting against fungal GH11 and GH12 glycoside hydrolase. However these proteins lack inhibitory activity. Here we describe the conversion of lupin γ-conglutin to an active inhibitor of endo-β-glucanases belonging to GH11 family. A set of γ-conglutin mutants was designed and expressed in Pichia pastoris, along with the wild-type protein. Unexpectedly, this latter was able to inhibit a GH11 enzyme, but not GH12, whereas the mutants were able to modulate the inhibition capacity. In lupin, γ-conglutin is naturally cleaved in two subunits, whereas in P. pastoris it is not. The lack of proteolytic cleavage is one of the reasons at the basis of the inhibitory activity of recombinant γ-conglutin. The results provide new insights about structural features at the basis of the lack of inhibitory activity of wild-type γ-conglutin and its legume homologues.
Seed proteins; Enzyme inhibitors; Mutagenesis; Plant defence
Settore BIO/10 - Biochimica
24-nov-2015
Article (author)
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/348825
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