Aims:
The aim of our study is to understand the biophysical traits that govern the stability and folding of Synechocystis hemoglobin, a unique cyanobacterial globin that displays unusual traits not observed in any of the other globins
discovered so far.
Background:
For the past few decades, classical hemoglobins such as vertebrate hemoglobin and myoglobin have been extensively studied to unravel the stability and folding mechanisms of hemoglobins. However, the expanding wealth of hemoglobins identified in all life forms with novel properties, like heme coordination chemistry and globin fold, have added
complexity and challenges to the understanding of hemoglobin stability, which has not been adequately addressed. Here, we
explored the unique truncated and hexacoordinate hemoglobin from the freshwater cyanobacterium Synechocystis sp. PCC
6803 known as “Synechocystis hemoglobin (SynHb)”. The “three histidines” linkages to heme are novel to this cyanobacterial hemoglobin.
Objective:
Mutational studies were employed to decipher the residues within the heme pocket that dictate the stability and
folding of SynHb.
Methods:
Site-directed mutants of SynHb were generated and analyzed using a repertoire of spectroscopic and calorimetric
tools. Result: The results revealed that the heme was stably associated to the protein under all denaturing conditions with
His117 playing the anchoring role. The studies also highlighted the possibility of existence of a “molten globule” like intermediate at acidic pH in this exceptionally thermostable globin. His117 and other key residues in the heme pocket play an
indispensable role in imparting significant polypeptide stability.
Conclusion:
Synechocystis hemoglobin presents an important model system for investigations of protein folding and stability in general. The heme pocket residues influenced the folding and stability of SynHb in a very subtle and specific manner
and may have been optimized to make this Hb the most stable known as of date.
Other:
The knowledge gained hereby about the influence of heme pocket amino acid side chains on stability and expression
is currently being utilized to improve the stability of recombinant human Hbs for efficient use as oxygen delivery vehicles.
Filling Up the Heme Pocket Stabilizes Apomyoglobin and Speeds Up Its Folding
