Aggregation-prone peptides modulate activity of bovine interferon gammareleased from naturally occurring protein nanoparticles

CARRATALÁA, J.V.; OLIVIA CANO-GARRIDO; SÁNCHEZ, JULIETA M; MEMBRADO, C.; PEREZ, E. ; CONCHILLO-SOLÉ, O.; DAURA, X.; SÁNCHEZ-CHARDI, ALEJANDRO; VILLAVERDE, ANTONIO; ARIS, A; GARCIA FRUITOS, E; FERRER MIRALLES, N

NEW BIOTECHNOLOGY

ELSEVIER SCIENCE BV

Lugar: Amsterdam; Año: 2019

1871-6784

Efficient protocols for the production of recombinant proteins are indispensable forthe development of the biopharmaceutical sector. Accumulation of recombinant proteins in naturally-occurring protein aggregates is detrimental to biopharmaceutical development.In recent years, the view of protein aggregates has completely changed with the recognition that these aggregates are a valuable source of functional recombinant proteins. In this study, bovine interferon-gamma (rBoIFN-)was engineered to enhance the formation of protein aggregates(also known as protein nanoparticles (NPs)) by the addition of aggregation-prone peptides(APPs) in the generally recognized as safe (GRAS) bacterialLactococcuslactis expression system. The L6K2, HALRU and CYOB peptides wereselected to assess theirintrinsic aggregation capability to nucleate protein aggregation. TheseAPPs enhanced the tendency to aggregate of the resulting protein at the expense of the total protein yield. However, fine physico-chemical characterization of the resulting intracellular protein NPs, the protein released from these protein NPs,and the protein purified from the soluble cell fractionindicated that the compactability of protein conformations is directly related to the biological activity of variants of IFN-, which is used here as a model protein with therapeutic potential.APPs enhance aggregation tendency of fused rBoIFN-while increasing compactability of protein species. Biological activity rBoIFN- is favored in more compacted conformations.Naturally-occurring protein aggregates can be produced in GRAS microorganisms as protein depots of releasable active protein. The addition of APPs to enhance aggregation tendency has a positive impact in overall compactability and functionality of resulting protein conformers.