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Preparation of recombinant rat eosinophil-associated ribonuclease-1 and -2 and analysis of their biological activities
- Source :
- Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease. 1638:164-172
- Publication Year :
- 2003
- Publisher :
- Elsevier BV, 2003.
-
Abstract
- 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.
- Subjects :
- Blood Bactericidal Activity
Hot Temperature
RNA, Transfer, Met
RNase P
Glycine
Saccharomyces cerevisiae
Eosinophil
Bactericidal activity
medicine.disease_cause
law.invention
Serine
Ribonucleases
law
Escherichia coli
otorhinolaryngologic diseases
medicine
Animals
Ribonuclease A activity
Amino Acid Sequence
Ribonuclease
Molecular Biology
Peptide sequence
chemistry.chemical_classification
biology
Granulocyte-Macrophage Colony-Stimulating Factor
Blood Proteins
Ribonuclease, Pancreatic
Eosinophil Granule Proteins
Molecular biology
Recombinant Proteins
Rats
Amino acid
Eosinophils
Kinetics
Biochemistry
chemistry
Recombinant DNA
biology.protein
Molecular Medicine
Cattle
sense organs
Subjects
Details
- ISSN :
- 09254439
- Volume :
- 1638
- Database :
- OpenAIRE
- Journal :
- Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease
- Accession number :
- edsair.doi.dedup.....82cae980037d0cd9d9cdaa5ecd7827e5