TY - CHAP
T1 - Quality Control of Proteins Solubilized from Inclusion Bodies
AU - Sánchez, Julieta M.
AU - Carratalá, Jose Vicente
AU - Gifre-Renom, Laia
AU - Arís, Anna
AU - Garcia-Fruitós, Elena
AU - Ferrer-Miralles, Neus
N1 - Publisher Copyright:
© 2022, Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2022
Y1 - 2022
N2 - Despite substantial development of production and purification protocols for heterologous recombinant proteins, some proteins are difficult to produce or, when produced, are accumulated in inclusion bodies (IBs). Nondenaturing protocols can be used to recover the entrapped protein from these protein aggregates. In this chapter, we provide a detailed procedure to analyze the physicochemical properties of one of those proteins produced in prokaryotic expression systems. Serum amyloid A3 (SAA3) was recovered from inclusion bodies (IBs) and its secondary structure associated to thermal stability and size was determined by circular dichroism (CD) and dynamic light scattering (DLS), respectively. These techniques were also applied to evaluate the SAA3 interaction with model membranes. These results show the importance of the structural analysis of proteins released from inclusion bodies under nondenaturing procedures, although similar approaches can be extended to any type of recombinant protein preparation.
AB - Despite substantial development of production and purification protocols for heterologous recombinant proteins, some proteins are difficult to produce or, when produced, are accumulated in inclusion bodies (IBs). Nondenaturing protocols can be used to recover the entrapped protein from these protein aggregates. In this chapter, we provide a detailed procedure to analyze the physicochemical properties of one of those proteins produced in prokaryotic expression systems. Serum amyloid A3 (SAA3) was recovered from inclusion bodies (IBs) and its secondary structure associated to thermal stability and size was determined by circular dichroism (CD) and dynamic light scattering (DLS), respectively. These techniques were also applied to evaluate the SAA3 interaction with model membranes. These results show the importance of the structural analysis of proteins released from inclusion bodies under nondenaturing procedures, although similar approaches can be extended to any type of recombinant protein preparation.
KW - Circular dichroism spectroscopy
KW - Dynamic light scattering
KW - Inclusion body
KW - Insoluble protein
KW - Multilamellar vesicles
KW - Recombinant protein
KW - Small unilamellar vesicles
KW - Thermal stability
UR - http://www.scopus.com/inward/record.url?scp=85123901464&partnerID=8YFLogxK
U2 - 10.1007/978-1-0716-1859-2_28
DO - 10.1007/978-1-0716-1859-2_28
M3 - Chapter
C2 - 35089575
AN - SCOPUS:85123901464
T3 - Methods in Molecular Biology
SP - 469
EP - 477
BT - Methods in Molecular Biology
ER -