Reminiscences: joyous moments along the road from here to there and back again

✍ Scribed by Allen B. Edmundson

• Publisher: John Wiley and Sons
• Year: 2002
• Tongue: English
• Weight: 153 KB
• Volume: 15
• Category: Article
• ISSN: 0952-3499
• DOI: 10.1002/jmr.583

✦ Synopsis

Sephadex was not commercially available for fractionation of biomolecules when I started graduate school at The Rockefeller University in 1955. Complete amino acid sequences were limited to the work of Fred Sanger (Nobel laureate) and his colleagues on the A (21 residues) and B (30 residues) chains of bovine insulin (Sanger and Tuppy 1951;Sanger 1952;Sanger and Thompson 1953;Sanger et al., 1955). The double helix model proposed for DNA by Watson and Crick (Watson and Crick 1953) was only two years old and the first X-ray analyses of proteins (myoglobin and hemoglobin) were just beginning to yield results in the laboratories of Kendrew, Perutz and Bragg (Kendrew et al., 1958;Perutz et al., 1960). My mentor, C.H.W. Hirs, was developing column chromatography methods for preparative-scale fractionation of peptides from the single polypeptide chain (124 residues) of oxidized ribonuclease A (Hirs 1956; Hirs et al., 1956a,b) and semi-quantitative procedures for determining their sequences by subtractive Edman degradation in solution.

These studies were being conducted in the laboratories of Stanford Moore and William Stein (1972 co-recipients of the Nobel Prize), whose great contribution was the first definitive procedure for quantitative amino acid analysis (Moore et al., 1958). Darrel Spackman, who had worked on aircraft engines prior to earning his Ph.D., was assembling the first automated amino acid analyzer with Dr Moore's daily help and guidance (Spackman et al., 1958;Spackman 1963). Dr Spackman also delineated the correct disulfide pairings of the eight half-cystine residues (105 possible combinations) in the native ribonuclease A molecule. By hydrolyzing the native form of ribonuclease A with pepsin and separating the peptides in acidic solutions, Dr Spackman avoided the disulfide interchange reactions, which had misled earlier investigators (Spackman et al., 1960).

After completing the preliminary reports on the sequence of oxidized ribonuclease A, Dr Hirs moved to Brookhaven National Laboratory, where he continued his brilliant work (Hirs 1960;Hirs et al., 1960). Dr Hirs was a gifted bio-and organic chemist, with a depth of instantly retrievable knowledge that I have never seen equaled. While technically remaining a Rockefeller student, I was allowed to accompany Dr Hirs to Brookhaven. When I returned to The Rockefeller University in 1960 to complete my thesis, Dr Hirs traveled 150 miles every Saturday from January to July to read, discuss, and correct every line and concept in the account of our joint work. His spirit has hovered over my shoulder for more than 45 years.