Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1976 Jun;73(6):1844–1847. doi: 10.1073/pnas.73.6.1844

Crystalline semisynthetic ribonuclease-S'.

M Pandin, E A Padlan, C DiBello, I M Chaiken
PMCID: PMC430403  PMID: 1064856

Abstract

Crystals of solid phase-derived semisynthetic ribonuclease-S' were prepared and compared with those for native ribonuclease-S' and -S. The semisynthetic species used was the noncovalent complex of synthetic fragment-(1-20), corresponding to residues 1 through 20 of bovine pancreatic ribonuclease-A (ribonucleate 3'-pyrimidino-oligonucleotidohydrolase, EC 3.1.4.22), and native ribonuclease-S-(21-124); the fragment containing residues 21 through 124 of ribonuclease-A. This semisynthetic complex was completely active enzymatically, was homogeneous as judged by polyacrylamide gel electrophoresis, and had no greater than trace amounts of excess ribonuclease-s(21-124) as judged by affinity chromatography. Crystallization of both semisynthetic and native ribonuclease-s' at pH 5.3 resulted in well-formed crystallseater than trace amounts of excess ribonuclease-S-T21-124) as judged by affinity chromatography. Crystallization of both semisynthetic and native ribonuclease-S' at pH 5.3 resulted in well-formed crystals with the symmetry of space group P3121 and unit cell dimensions a=b=44.82, c=97.3 A. This crystal form corresponds to the Y form of native ribonuclease-S previously reported [Wyckoff et al. (1967) J. Biol. Chem. 242, 3749-3753]. X-ray diffraction patterns of the crystals were indistinguishable, indicative of the structural identity of semisynthetic and native ribonuclease-S'.

Full text

PDF
1844

Images in this article

1845Image
on p.1845
1846Image
on p.1846
1846Image
on p.1846

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Chaiken I. M., Freedman M. H., Lyerla J. R., Jr, Cohen J. S. Preparation and studies of 19 F-labeled and enriched 13 C-labeled semisynthetic ribonuclease-S' analogues. J Biol Chem. 1973 Feb 10;248(3):884–891. [PubMed] [Google Scholar]
  2. Chaiken I. M. Purification and properties of semisynthetic staphylococcal nuclease-T'. J Biol Chem. 1971 May 10;246(9):2948–2952. [PubMed] [Google Scholar]
  3. Doscher M. S., Hirs C. H. The heterogeneity of bovine pancreatic ribonuclease S. Biochemistry. 1967 Jan;6(1):304–312. doi: 10.1021/bi00853a047. [DOI] [PubMed] [Google Scholar]
  4. Dunn B. M., Chaiken I. M. Relationship between alpha-helical propensity and formation of the ribonuclease-S complex. J Mol Biol. 1975 Jul 15;95(4):497–511. doi: 10.1016/0022-2836(75)90313-7. [DOI] [PubMed] [Google Scholar]
  5. Dunn B. M., DiBello C., Kirk K. L., Cohen L. A., Chaiken I. M. Synthesis, purification, and properties of a semisynthetic ribonuclease S incorporating 4-fluoro-L-histidine at position 12. J Biol Chem. 1974 Oct 10;249(19):6295–6301. [PubMed] [Google Scholar]
  6. Lin M. C., Gutte B., Moore S., Merrifield R. B. Regeneration of activity by mixture of ribonuclease enzymically degraded from the COOH terminus and a synthetic COOH-terminal tetradecapeptide. J Biol Chem. 1970 Oct 10;245(19):5169–5170. [PubMed] [Google Scholar]
  7. Merrifield R. B. Automated synthesis of peptides. Science. 1965 Oct 8;150(3693):178–185. doi: 10.1126/science.150.3693.178. [DOI] [PubMed] [Google Scholar]
  8. Ontjes D. A., Anfinsen C. B. Solid phase synthesis of a 42-residue fragment of staphylococcal nuclease: properties of a semisynthetic enzyme. Proc Natl Acad Sci U S A. 1969 Oct;64(2):428–435. doi: 10.1073/pnas.64.2.428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Parikh I., Corley L., Anfinsen C. B. Semisynthetic analogues of an enzymically active complex formed between two overlapping fragments of staphylococcal nuclease. J Biol Chem. 1971 Dec 10;246(23):7392–7397. [PubMed] [Google Scholar]
  10. REISFELD R. A., LEWIS U. J., WILLIAMS D. E. Disk electrophoresis of basic proteins and peptides on polyacrylamide gels. Nature. 1962 Jul 21;195:281–283. doi: 10.1038/195281a0. [DOI] [PubMed] [Google Scholar]
  11. RICHARDS F. M., VITHAYATHIL P. J. The preparation of subtilisn-modified ribonuclease and the separation of the peptide and protein components. J Biol Chem. 1959 Jun;234(6):1459–1465. [PubMed] [Google Scholar]
  12. Rocchi R., Moroder L., Marchiori F., Ferrarese E., Scoffone E. Synthesis of peptide analogs of the N-terminal eicosapeptide sequence of ribonuclease A. 8. Synthesis of (ser4, orn10)-and (ser5, orn10)-eicosapeptides. J Am Chem Soc. 1968 Oct 9;90(21):5885–5889. doi: 10.1021/ja01023a039. [DOI] [PubMed] [Google Scholar]
  13. Wyckoff H. W., Hardman K. D., Allewell N. M., Inagami T., Tsernoglou D., Johnson L. N., Richards F. M. The structure of ribonuclease-S at 6 A resolution. J Biol Chem. 1967 Aug 25;242(16):3749–3753. [PubMed] [Google Scholar]
  14. Wyckoff H. W., Tsernoglou D., Hanson A. W., Knox J. R., Lee B., Richards F. M. The three-dimensional structure of ribonuclease-S. Interpretation of an electron density map at a nominal resolution of 2 A. J Biol Chem. 1970 Jan 25;245(2):305–328. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES