Open AccessMolten globule as an intermediate on the human prostatic phosphatase folding pathway.
Author(s) -
Radosława Kuciel,
Aleksandra Mazurkiewicz
Publication year - 1997
Publication title -
acta biochimica polonica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.452
H-Index - 78
eISSN - 1734-154X
pISSN - 0001-527X
DOI - 10.18388/abp.1997_4367
Subject(s) - molten globule , chemistry , circular dichroism , folding (dsp implementation) , liposome , protein folding , monomer , biophysics , amphiphile , enzyme , phosphatidylserine , phospholipid , biochemistry , organic chemistry , polymer , membrane , electrical engineering , copolymer , biology , engineering
Human prostatic acid phosphatase (hPAP, EC.3.1.3.2), a secretory homodimeric protein was denatured in 6 M urea, pH 2.5, and refolded by dilution at pH 7.2 with recovery of the enzymatic activity and dimeric structure. Circular dichroism, intrinsic fluorescence and chromatographic analysis of renaturating protein suggested that the kinetic intermediate of the hPAP folding is a monomer which displays a molten globule state (R. Kuciel, A. Mazurkiewicz & W.S. Ostrowski, 1996, Int. J. Biol. Macromol. 18, 167-175). To confirm these data experiments were performed to estimate the interaction of the renaturating protein with dyes and amphipathic lipid structures. Increased binding of the hydrophobic probe 1-anilinonaphthalene-8-sulfonate and Congo Red to the refolding enzyme supported the existence of molten globule state with the relaxed beta-structure in the renaturating protein. Presence of liposomes, included in the renaturation mixture as a model of acid phospholipid, resulted in perturbations of the human PAP refolding process. Some folding intermediates were bound to phosphatidylserine liposomes or, alternatively, water soluble, inactive aggregates were formed.