Document Detail

Preparation of recombinant rat eosinophil-associated ribonuclease-1 and -2 and analysis of their biological activities.
MedLine Citation:
PMID:  12853122     Owner:  NLM     Status:  MEDLINE    
Rat eosinophils contain eosinophil-associated ribonucleases (Ears) in their granules. Ears are thought to be synthesized as pre-forms and stored in the granules as mature forms. However, the N-terminal amino acid of mature Ear-1 and Ear-2 is still controversial. Therefore, we prepared two recombinant mature forms of Ear-1 and Ear-2 in which the N-terminal amino acids are Ser24 (S) [Ear-1 (S) and Ear-2 (S)] and Gln26 (Q) [Ear-1 (Q) and Ear-2 (Q)], and analyzed their biological activities by comparing them with those of pre-form Ear-1 and pre-form Ear-2. The four mature Ears showed RNase A activity as well as bovine pancreatic RNase A activity, but pre-Ear-1 and pre-Ear-2 showed no RNase A activity. Mature Ear-1 (Q) and mature Ear-2 (Q) showed more potent RNase A activity than mature Ear-1 (S) and mature Ear-2 (S), respectively. The RNase A activities of mature Ear-1 (Q) and mature Ear-2 (Q) were reduced by treatment at 96 degrees C for 20 min or with RNase inhibitor. The growth of Escherichia coli was inhibited by both pre-Ears and mature Ears in a concentration-dependent manner, and was almost completely suppressed at 1.0 microM. The bactericidal activities of mature Ear-1 (Q) and mature Ear-2 (Q) were not inhibited by RNase inhibitor, but was increased by treatment at 96 degrees C for 20 min.
Kenji Ishihara; Kanako Asai; Masahiro Nakajima; Suetsugu Mue; Kazuo Ohuchi
Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biochimica et biophysica acta     Volume:  1638     ISSN:  0006-3002     ISO Abbreviation:  Biochim. Biophys. Acta     Publication Date:  2003 Jul 
Date Detail:
Created Date:  2003-07-10     Completed Date:  2003-08-21     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  0217513     Medline TA:  Biochim Biophys Acta     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  164-72     Citation Subset:  IM    
Laboratory of Pathophysiological Biochemistry, Graduate School of Pharmaceutical Sciences, Tohoku University, Aoba Aramaki, Aoba-ku, Sendai, Miyagi 980-8578, Japan.
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MeSH Terms
Amino Acid Sequence
Blood Bactericidal Activity
Blood Proteins
Eosinophil Granule Proteins
Eosinophils / enzymology,  metabolism*
Escherichia coli
Glycine / metabolism
Granulocyte-Macrophage Colony-Stimulating Factor / metabolism
Hot Temperature
RNA, Transfer, Met / metabolism
Recombinant Proteins / chemistry,  isolation & purification,  metabolism*
Ribonuclease, Pancreatic / blood,  drug effects,  genetics,  metabolism
Saccharomyces cerevisiae
Serine / metabolism
Reg. No./Substance:
0/Blood Proteins; 0/Eosinophil Granule Proteins; 0/RNA, Transfer, Met; 0/Recombinant Proteins; 56-40-6/Glycine; 56-45-1/Serine; 83869-56-1/Granulocyte-Macrophage Colony-Stimulating Factor; EC 3.1.-/Ribonucleases; EC, Pancreatic

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