The structural basis of the thermostability of SP1, a novel plant (Populus tremula) boiling stable protein

Or Dgany, Ana Gonzalez, Oshrat Sofer, Wangxia Wang, Gennady Zolotnitsky, Amnon Wolf, Yuval Shoham, Arie Altman, Sharon G. Wolf, Oded Shoseyov, Orna Almog*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

71 Scopus citations

Abstract

We previously reported on a new boiling stable protein isolated from aspen plants (Populus tremula), which we named SP1. SP1 is a stress-related protein with no significant sequence homology to other stress-related proteins. It is a 108-amino-acid hydrophilic polypeptide with a molecular mass of 12.4 kDa (Wang, W. X., Pelah, D., Alergand, T., Shoseyov, O., and Altman, A. (2002) Plant Physiol. 130, 865-875) and is found in an oligomeric form. Preliminary electron microscopy studies and matrix-assisted laser desorption ionization time-of-fliglit mass spectrometry experiments showed that SP1 is a dodecamer composed of two stacking hexamers. We performed a SDS-PAGE analysis, a differential scanning calorimetric study, and crystal structure determination to further characterize SP1. SDS-PAGE indicated a spontaneous assembly of SP1 to one stable oligomeric form, a dodecamer. Differential scanning calorimetric showed that SP1 has high thermostability i.e. Tm of 107°C (at pH 7.8). The crystal structure of SP1 was initially determined to 2.4 Å resolution by multi-wave-length anomalous dispersion method from a crystal belonging to the space group 1422. The phases were extended to 1.8 Å resolution using data from a different crystal form (P21). The final refined molecule includes 106 of the 108 residues and 132 water molecules (on average for each chain). The R-free is 20.1%. The crystal structure indicated that the SP1 molecule has a ferredoxin-like fold. Strong interactions between each two molecules create a stable dimer. Six dimers associate to form a ring-like-shaped dodecamer strongly resembling the particle visualized in the electron microscopy studies. No structural similarity was found between the crystal structure of SP1 and the crystal structure of other stress-related proteins such as small heat shock proteins, whose structure has been already determined. This structural study further supports our previous report that SP1 may represent a new family of stress-related proteins with high thermostability and oligomerization.

Original languageAmerican English
Pages (from-to)51516-51523
Number of pages8
JournalJournal of Biological Chemistry
Volume279
Issue number49
DOIs
StatePublished - 3 Dec 2004

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