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Nucleic Acids Research, 2000, Vol. 28, No. 16 3075-3082
© 2000 Oxford University Press

Proteomics of Mycoplasma genitalium: identification and characterization of unannotated and atypical proteins in a small model genome

Suganthi Balasubramanian1, Tamara Schneider1, Mark Gerstein1,2 and Lynne Regan1,3,*

1Department of Molecular Biophysics and Biochemistry, 2Department of Computer Science and 3Department of Chemistry, Yale University, 266 Whitney Avenue, New Haven, CT 06520-8114, USA

We present the results of a comprehensive analysis of the proteome of Mycoplasma genitalium (MG), the smallest autonomously replicating organism that has been completely sequenced. Our aim was to identify and characterize all soluble proteins in MG that are structurally and functionally uncharacterized. We were particularly interested in identifying proteins that differed significantly from typical globular proteins, for example, proteins which are unstructured in the absence of a ‘partner’ molecule or those that exhibit unusual thermodynamic properties. This work is complementary to other structural genomics projects whose primary aim is to determine the three-dimensional structures of proteins with unknown folds. We have identified all the full-length open reading frames (ORFs) in MG that have no homologs of known structure and are of unknown function. Twenty-five of the total 483 ORFs fall into this category and we have expressed, purified and characterized 11 of them. We have used circular dichroism (CD) to rapidly investigate their biophysical properties. Our studies reveal that these proteins have a wide range of structures varying from highly helical to partially structured to unfolded or random coil. They also display a variety of thermodynamic properties ranging from cooperative unfolding to no detectable unfolding upon thermal denaturation. Several of these proteins are highly conserved from mycoplasma to man. Further information about target selection and CD results is available at http://bioinfo.mbb.yale.edu/genome

* To whom correspondence should be addressed. Tel: +1 203 432 9843; Fax: +1 203 432 5175; Email: lynne@csb.yale.edu


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