Abstract
A non-specific protease, subtilisin, was used to probe the existence of folded structure in thermal aggregates of bovine carbonic anhydrase-II (BCA). BCA aggregates and native BCA were subjected to proteolysis and electrophoretic analyses which revealed the accumulation of polypeptide fragments of similar size, indicating survival of similar sections of folded structure burying peptide bonds away from scission in the two samples. N-terminal sequencing revealed that the termini of size-matched fragments from the two samples were either identical, or located very close to each other, and predominantly on the surface of the 3-dimensional structure of native BCA. The susceptibility to proteolysis of very nearly the same sites in the two samples suggests that native-like elements of structure survive within BCA aggregates. The finding that thermal aggregation can involve interactions among molecules retaining elements of native-like structure, suggests that complete chain unfolding may not be a necessary prerequisite for all aggregation.
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Acknowledgements
SS thanks the Council of Scientific & Industrial Research (CSIR), India, for a doctoral research fellowship. PG thanks the Indian National Science Academy (INSA) and the CSIR, for grants supporting research into protein folding and aggregation.
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Sharma, S., Guptasarma, P. Evidence of Native-like Substructure(s) in Polypeptide Chains of Carbonic Anhydrase Deposited into Insoluble Aggregates During Thermal Unfolding. Protein J 27, 50–58 (2008). https://doi.org/10.1007/s10930-007-9107-y
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DOI: https://doi.org/10.1007/s10930-007-9107-y