Structure, stability, and orientation of BSA adsorbed to silica.
In this investigation, the structure, stability, and orientation of bovine serum albumin (BSA) adsorbed onto silica particles were studied using differential scanning calorimetry (DSC) and limited proteolysis in combination with mass spectrometry (MS). DSC gave information on the overall structural stability of BSA while limited proteolysis was used to probe the accessibility of enzymatic cleavage sites, thereby yielding information on the orientation and structure of BSA adsorbed to silica surfaces. Thermal investigation of BSA in various buffers, both free in solution and in the adsorbed state, showed that solutes that surround the protein played an important role with respect to the overall structural stability and the structural heterogeneity of BSA. Limited proteolysis with trypsin and chymotrypsin indicated that BSA in the adsorbed state is oriented with domain 2 facing the silica surface. Also, upon adsorption, no additional cleavage sites were exposed. The combination of the results presented in this study implied that BSA molecules adsorbed onto silica particles were significantly reduced in their structural stability, but not to an extent that internal residues within the native structure became fully exposed to the solution.
Larsericsdotter
Helén
H
Department of Chemical Engineering, Mälardalen University, Eskilstuna, Sweden. helen.larsericsdotter@mdh.se
Oscarsson
Sven
S
Buijs
Jos
J
D000327:Adsorption; D000595:Amino Acid Sequence; D000818:Animals; D002152:Calorimetry, Differential Scanning / Q000379:methods; D002417:Cattle; D002918:Chymotrypsin / Q000737:chemistry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D012822:Silicon Dioxide / Q000737:chemistry*; D013329:Structure-Activity Relationship; D013499:Surface Properties; D013696:Temperature; D014357:Trypsin / Q000737:chemistry
D012710:Serum Albumin, Bovine; D012822:Silicon Dioxide; D002918:Chymotrypsin; D014357:Trypsin