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Amino acids and peptides. LVII. Synthetic peptide with a sequence of ribonuclease from Sulfolobus solfataricus , SSR(1–62), does not function as an RNase
Author(s) -
Joshi Shima,
Tsuda Yuko,
Shintomi Noriyuki,
Kondo Hiroyuki,
Nishiyama Yasuhiro,
Iwama Masanori,
Ohgi Kazuko,
Irie Masachika,
Okada Yoshio
Publication year - 2000
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(00)01185-6
Subject(s) - sulfolobus solfataricus , rnase p , biochemistry , biology , dna , peptide , rnase h , ribonuclease , transfer rna , restriction enzyme , microbiology and biotechnology , rna , chemistry , gene , archaea
The 62 residue peptide, SSR(1–62), whose sequence corresponds to that of ribonuclease (RNase) from Sulfolobus solfataricus , and its related peptides, SSR(1–22) and SSR(10–62), were chemically synthesized and their RNase activity and DNA‐binding activity were examined. The RNase activity assay using yeast RNA or t RNA fMet as substrate showed that the synthetic peptide SSR(1–62) did not hydrolyze yeast RNA or t RNA fMet . These data were not consistent with previous reports that both the native peptide isolated from S. solfataricus [Fusi et al. (1993) Eur. J. Biochem. 211, 305–311] and the recombinant peptide expressed in Escherichia coli [Fusi et al. (1995) Gene 154, 99–103] were able to hydrolyze t RNA fMet . However, the synthetic SSR(1–62) exhibited DNA‐binding activity. In the presence of synthetic SSR(1–62), the cleavage of DNA (plasmid pUCRh2‐4) by restriction endonuclease ( Eco RI) was not observed, suggesting that synthetic SSR(1–62) bound to DNA protected DNA from its enzymatic digestion. Neither SSR(1–22) nor SSR(10–62) prevented DNA from being cleaved by a restriction enzyme. These findings strongly suggest the importance of not only the N‐terminal region of SSR(1–62) but also the C‐terminal region for DNA‐binding. Circular dichroism spectroscopy of synthetic SSR(1–62) indicated a β‐sheet conformation, in contrast with synthetic SSR(1–22), which exhibited an unordered conformation.