Ribonuclease activity and substrate preference of human eosinophil cationic protein (ECP)

Sorrentino, Salvatore; Glitz, Dohn G.

doi:10.1016/0014-5793(91)80994-e

Cited by 36 publications

(38 citation statements)

References 29 publications

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“…Early kinetic studies on the RNase activity of ECP indicated a pyrimidine specificity for the base position at the active site with a slight preference for cytidine in substrates of low mo-lecular mass but a clear preference for polyuridylic acid (poly(U)), in comparison with polycytidylic acid (poly(C)) (18,19). No activity toward either 2Ј,3Ј-phosphate cyclic mononucleotides, dinucleoside monophosphates, or double-stranded RNA was detected (20).…”

Section: Human Eosinophil Cationic Protein (Ecp)mentioning

confidence: 99%

Kinetic and Product Distribution Analysis of Human Eosinophil Cationic Protein Indicates a Subsite Arrangement That Favors Exonuclease-type Activity

Boix¹,

Николовски²,

Moiseyev³

et al. 1999

Journal of Biological Chemistry

101

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With the use of a high yield prokaryotic expression system, large amounts of human eosinophil cationic protein (ECP) have been obtained. This has allowed a thorough kinetic study of the ribonuclease activity of this protein. The catalytic efficiencies for oligouridylic acids of the type (Up) n U>p, mononucleotides U>p and C>p, and dinucleoside monophosphates CpA, UpA, and UpG have been interpreted by the specific subsites distribution in ECP. The distribution of products derived from digestion of high molecular mass substrates, such as poly(U) and poly(C), by ECP was compared with that of RNase A. The characteristic cleavage pattern of polynucleotides by ECP suggests that an exonucleaselike mechanism is predominantly favored in comparison to the endonuclease catalytic mechanism of RNase A. Comparative molecular modeling with bovine pancreatic RNase A-substrate analog crystal complexes revealed important differences in the subsite structure, whereas the secondary phosphate-binding site (p 2 ) is lacking, the secondary base subsite (B 2 ) is severely impaired, and there are new interactions at the p o , B o , and p -1 sites, located upstream of the P-O-5 cleavable phosphodiester bond, that are not found in RNase A. The differences in the multisubsites structure could explain the reduced catalytic efficiency of ECP and the shift from an endonuclease to an exonuclease-type mechanism.

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Section: Human Eosinophil Cationic Protein (Ecp)mentioning

confidence: 99%

Kinetic and Product Distribution Analysis of Human Eosinophil Cationic Protein Indicates a Subsite Arrangement That Favors Exonuclease-type Activity

Boix¹,

Николовски²,

Moiseyev³

et al. 1999

Journal of Biological Chemistry

101

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“…This protein potently elicits neovascularization (Fett et al, 1985;Strydom et al, 1985), induccs second-messenger responses in cultured endothelial cells Vallee, 1988, 1989), and interacts with putative cell-surface receptors (Hu et al, 1991;Badet et al, 1989;Chainoux et al, 1991 ). Eosinophil-derived neurotoxin (EDN) and eosinophil cationic protein (ECP) are also ribonucleases, homologous to pancreatic RNase Beintema et al, 1988b;Rosenberg et al, 1989b;Hamann et al, 1989;Barker el al., 1989;Slifman et al, 3986;Gullberg et al, 1986;Sorrentino and Glitz, 1991), that in this case act as neurotoxins and/or helminthotoxins (Durack et al, 1981 ;Fredens et al, 1982;McLaren et a]., 1984;Sorrentino et al, 1992). It is now apparent through these and other studies (Beintema et al, 1988b) that the human RNases with known sequences can be classified into four types as follows: (a) secretory/ pancreatic, (b) the so-called non-secretory (EDN), (c) ECP and (d) angiogenin.…”

mentioning

confidence: 99%

The amino acid sequence of human ribonuclease 4, a highly conserved ribonuclease that cleaves specifically on the 3′ side of uridine

Zhou

Strydom

1993

European Journal of Biochemistry

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. (1986a) Biochemistry 25,7255-72641 was purified from human plasma and its amino acid sequence was determined. This protein is a 119-residue single-chain polypeptidc cross-linked by four disulfide bonds and has an amino-terminal pyroglutaminyl residue. No post-translational modifications were observed during extensive sequence studies on peptide fragments, except for the amino-terminal pyroglutamic acid and a possible deamidation of Asn66. The protein is homologous to the pancreatic ribonucleases and angiogenin, but differs substantially from both of these proteins; the protein sequence has 43% identity with human pancreatic ribonuclease and 39% identity with human angiogenin, as compared to 35% identity between human angiogenin and pancreatic ribonuclease. It is referred to as RNase 4, based on the nomenclature currently used for the genes of pancreatic RNase (RNase 1) and the eosinophil-derived RNases (RNase 2 and RNase 3).Virtually all of the RNasc active-site componcnts, including the catalytic residues Hisl2, His1 19 and Lys41, are preserved. However, some invariant residues of RNase 1 arc replaced, e.g. Lys7 by arginine, Asp14 by histidine, and Pro42 by arginine. RNase 4 contains a unique two-residue deletion at the position corresponding to amino acids 77 and 78 of pancreatic RNase, and its carboxyterminal sequence is truncated at position 122. The deletion in angiogenin at position 21 is also found in RNase 4.RNase 4 is very similar to two RNases isolated from bovine and porcine liver, and together they form a new fanlily in the RNase superfamily. The degree of inter-species similarity (90%) is much greater than within the pancreatic RNase and angiogenin families, which suggests that this ribonuclease could possess a physiologically important function other than general RNA catabolism.The ribonucleases (RNases) constitute a group of enzymes for which a large amount of chemical information now exists. This information mostly relates to the pancreatic RNases; sequences from more than 40 species are known (Beintema and van der Laan, 1986). Recent investigations and discoveries have suggested that these enzymes are but one subdivision of a much broader superfamily of structurally related proteins, some of which have important biochemical and physiological functions other than RNA digestion.One homologue of pancreatic RNase that has an unusual ribonucleolytic activity is angiogenin (Shapiro et al., 1986b). This protein potently elicits neovascularization (Fett et al., 1985; Strydom et al., 1985), induccs second-messenger responses in cultured endothelial cells (Bicknell and Vallee, Con-rsyondeirce ro D. J. Strydom,

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“…These granule proteins appear to provide effector functions independent of their associated ribonuclease activities. ECP is toxic to mammalian cells and has demonstrable bactericidal and anti-helminthic activities (19)(20)(21)(22)(23), yet its associated ribonuclease activity is relatively weak (15,24). In contrast, EDN has little generalized toxicity (19,20) but is an efficient ribonuclease having activity similar to pancreatic RNase A (24)(25)(26).…”

mentioning

confidence: 99%

Two highly homologous ribonuclease genes expressed in mouse eosinophils identify a larger subgroup of the mammalian ribonuclease superfamily.

Larson

Olson

Madden

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

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Two putative ribonucleases have been isolated from the secondary granules of mouse eosinophils. Degenerate oligonucleotide primers inferred from peptide sequence data were used in reverse transcriptase-PCR reactions of bone marrow-derived cDNA. The resulting PCR product was used to screen a C57BL/6J bone marrow cDNA library, and comparisons of representative clones showed that these genes and encoded proteins are highly homologous (96% identity at the nucleotide level; 92/94% identical/similar at the amino acid level). The mouse proteins are only weakly homologous (-50%o amino acid identity) with the human eosinophil-associated ribonucleases (i.e., eosinophil-derived neurotoxin and eosinophil cationic protein) and show no sequence bias toward either human protein. Phylogenetic

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Ribonuclease activity and substrate preference of human eosinophil cationic protein (ECP)

Cited by 36 publications

References 29 publications

Kinetic and Product Distribution Analysis of Human Eosinophil Cationic Protein Indicates a Subsite Arrangement That Favors Exonuclease-type Activity

Kinetic and Product Distribution Analysis of Human Eosinophil Cationic Protein Indicates a Subsite Arrangement That Favors Exonuclease-type Activity

The amino acid sequence of human ribonuclease 4, a highly conserved ribonuclease that cleaves specifically on the 3′ side of uridine

Two highly homologous ribonuclease genes expressed in mouse eosinophils identify a larger subgroup of the mammalian ribonuclease superfamily.

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