Back to the future: Ribonuclease A

Abstract

Pancreatic ribonuclease A (EC 3.1.27.5, RNase) is, perhaps, the best‐studied enzyme of the 20th century. It was isolated by René Dubos, crystallized by Moses Kunitz, sequenced by Stanford Moore and William Stein, and synthesized in the laboratory of Bruce Merrifield, all at the Rockefeller Institute/University. It has proven to be an excellent model system for many different types of experiments, both as an enzyme and as a well‐characterized protein for biophysical studies. Of major significance was the demonstration by Chris Anfinsen at NIH that the primary sequence of RNase encoded the three‐dimensional structure of the enzyme. Many other prominent protein chemists/enzymologists have utilized RNase as a dominant theme in their research. In this review, the history of RNase and its offspring, RNase S (S‐protein/S‐peptide), will be considered, especially the work in the Merrifield group, as a preface to preliminary data and proposed experiments addressing topics of current interest. These include entropy–enthalpy compensation, entropy of ligand binding, the impact of protein modification on thermal stability, and the role of protein dynamics in enzyme action. In continuing to use RNase as a prototypical enzyme, we stand on the shoulders of the giants of protein chemistry to survey the future. © 2007 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 90: 259–277, 2008.This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com

Electronic and vibrational signatures of peptide helical structures: A tribute to Anton Mario Tamburro

“…1), an helicogenic C a -tetrasubstituted a-amino acid of the Aib family. 43 In addition, a more detailed FTIR absorption investigation on the -(Aib) 8 -homo-oligomer in the 1800-1500 cm 21 region, in combination with its deconvolved spectra and second-derivative spectra (Fig. 9A), showed that the amide-I and amide-II bands for this 3 10 -helical peptide occur at 1666 and 1532 cm 21 , respectively.…”

Genotypes and Phenotypes in the Evolution of Molecules

“…Historically, the concept that protein folding follows a straightforward and reversible downhill process and ends at the thermodynamically stable (and therefore uniquely characterized) conformation of the molecule was derived from early work on the protein bovine pancreatic ribonuclease A (Raines, 1998;Marshall et al, 2008). The sequence of the small protein with a chain length of 124 amino acids was determined by Stanford Moore and William Stein (Smyth et al, 1963;Moore and Stein, 1973) and only 4 years later the threedimensional molecular structure of the protein had been determined (Kartha et al, 1967;Wyckoff et al, 1967aWyckoff et al, , 1967bKim et al, 1992).…”

Incorporation of non-natural modules into proteins: structural features beyond the genetic code

“…The finally resulting alloprotein, which displayed enzymatic activity like RNase A and was slightly more resistant to thermal denaturation (Arnold et al 2002a), was the first protein with altered backbone structure in its interior generated by use of EPL. However, the two additional methylene groups in the rather bulky mimetic (for a structural alignment and an excellent analysis of bond lengths and dihedral angles, see Marshall et al 2008) might prevent a more significant stabilization of RNase A by impairing the native fold. Thus, we replaced Pro114 with 5,5 0 -dimethylproline (dmP, Fig.…”