Properties of truncated forms of the elongation factor 1alpha from the archaeon Sulfolobus solfataricus.
Two truncated forms of the Sulfolobus solfataricus elongation factor 1alpha (SsEF-1alpha), corresponding to the putative domains G+M, Ss(GM)EF-1alpha, and G, Ss(G)EF-1alpha, have been constructed by gene engineering, produced in Escherichia coli and purified. Neither truncated form was able to sustain poly(Phe) synthesis but they were able to bind guanine nucleotides with an affinity much higher with respect to that of the intact factor. However, the difference in the affinity for GDP and GTP became progressively reduced with the extent of the truncation. The values of kcat and Km for GTP of the intrinsic GTPase of SsEF-1alpha triggered by 3.6 M NaCl were not affected by the deletions. In contrast, both Ss(GM)EF-1alpha and Ss(G)EF-1alpha were less thermostable than the intact factor; the region of the factor most responsible for the loss of resistance against heat inactivation was the C-terminal domain. On the other hand the domain M was the regulator of the thermophilicity of SsEF-1alpha since only Ss(G)EF-1alpha showed a reduced thermophilicity. Remarkably, both Ss(GM)EF-1alpha and Ss(G)EF-1alpha were able to exchange [3H]GDP for GTP at a very high rate so that they were no more sensitive to the stimulatory effect of SsEF-1beta, which is the nucleotide exchange factor of SsEF-1alpha.
Masullo
M
M
Dipartimento di Biochimica e Biotecnologie Mediche, Università di Napoli Federico II, Italy.
Ianniciello
G
G
Arcari
P
P
Bocchini
V
V
D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D019277:Entropy; D020963:GTP Phosphohydrolase-Linked Elongation Factors / Q000378:metabolism; D006150:Guanine Nucleotides / Q000378:metabolism; D006358:Hot Temperature; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors / Q000737:chemistry / Q000378:metabolism*; D014176:Protein Biosynthesis; D011489:Protein Denaturation; D017384:Sequence Deletion; D013329:Structure-Activity Relationship; D016944:Sulfolobus / Q000378:metabolism*; D013816:Thermodynamics
D001426:Bacterial Proteins; D006150:Guanine Nucleotides; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors; D020963:GTP Phosphohydrolase-Linked Elongation Factors