TY - JOUR
T1 - Role of the intersubunit disulfide bond in the unfolding pathway of dimeric red kidney bean purple acid phosphatase
AU - Cashikar, Anil G.
AU - Rao, N. Madhusudhana
N1 - Funding Information:
G.A.C. was a recipient of the Senior Research Fellowship from the Council for Scientific and Industrial Research (CSIR), Government of India.
PY - 1996/8/15
Y1 - 1996/8/15
N2 - Quantitative equilibrium denaturation studies on oligomeric proteins have the potential to provide information on the role of subunit interactions in protein function and structure. We studied the equilibrium denaturation of red kidney bean purple acid phosphatase (KBPAP), a homodimer with a single disulfide bond between the two subunits, with an objective to understand the role of the intersubunit disulfide bond in KBPAP structure. Binding of 8-anilino-1-naphthalenesulfonic acid, enzymatic activity, size-exclusion chromatography, tryptophan fluorescence and circular dichroism studies revealed that the protein undergoes unfolding through at least three intermediates. Susceptibility of KBPAP for denaturation increases on reduction of the disulfide and aggregation was the predominant product of denaturation. In terms of stability, an intersubunit disulfide bond contributes to 25% of the overall stability of the dimer.
AB - Quantitative equilibrium denaturation studies on oligomeric proteins have the potential to provide information on the role of subunit interactions in protein function and structure. We studied the equilibrium denaturation of red kidney bean purple acid phosphatase (KBPAP), a homodimer with a single disulfide bond between the two subunits, with an objective to understand the role of the intersubunit disulfide bond in KBPAP structure. Binding of 8-anilino-1-naphthalenesulfonic acid, enzymatic activity, size-exclusion chromatography, tryptophan fluorescence and circular dichroism studies revealed that the protein undergoes unfolding through at least three intermediates. Susceptibility of KBPAP for denaturation increases on reduction of the disulfide and aggregation was the predominant product of denaturation. In terms of stability, an intersubunit disulfide bond contributes to 25% of the overall stability of the dimer.
KW - Denaturation
KW - Intersubunit disulfide bond
KW - Phosphatase
KW - Purple acid phosphatase
KW - Red kidney bean
KW - Unfolding intermediate
UR - http://www.scopus.com/inward/record.url?scp=0030586786&partnerID=8YFLogxK
U2 - 10.1016/0167-4838(96)00055-6
DO - 10.1016/0167-4838(96)00055-6
M3 - Article
C2 - 8765232
AN - SCOPUS:0030586786
SN - 0167-4838
VL - 1296
SP - 76
EP - 84
JO - Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology
JF - Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology
IS - 1
ER -