Ninety-eight semesters of cytochrome P450 enzymes and related topics-What have I taught and learned?

Abstract

This Reflection article begins with my family background and traces my career through elementary and high school, followed by time at the University of Illinois, Vanderbilt University, the University of Michigan, and then for 98 semesters as a Vanderbilt University faculty member. My research career has dealt with aspects of cytochrome P450 enzymes, and the basic biochemistry has had applications in fields as diverse as drug metabolism, toxicology, medicinal chemistry, pharmacogenetics, biological engineering, and bioremediation. I am grateful for the opportunity to work with the Journal of Biological Chemistry not only as an author but also for 34 years as an Editorial Board Member, Associate Editor, Deputy Editor, and interim Editor-in-Chief. Thanks are extended to my family and my mentors, particularly Profs. Harry Broquist and Minor J. Coon, and the more than 170 people who have trained with me. I have never lost the enthusiasm for research that I learned in the summer of 1968 with Harry Broquist, and I have tried to instill this in the many trainees I have worked with. A sentence I use on closing slides is "It's not just a laboratory-it's a fraternity."

Keywords: cytochrome P450; electron transfer; enzyme kinetics; enzyme mechanisms; mentoring; oxidation-reduction; steroid metabolism; toxicology drug metabolism.

Figure 1

The author at work driving a corn picker (ca. age 5). As mentioned, my father had me working on the farm at a young age, but not quite yet—this shot was staged. I did not learn to drive that tractor (Farmall Model H) until the age of 10. (Incidentally, corn pickers have been replaced today by combines.)

Figure 2

The author in college (early 1970). The instrument is a Beckman Model DU spectrophotometer (popular then), with a centrifuge rotor in the foreground.

Figure 3

The author (left) with Ph.D. thesis mentor Harry Broquist (right).This wasin 2007 on the occasion of receiving an endowed chair at Vanderbilt.

Figure 4

The author’s postdoctoral mentor Jud Coon (left) presenting him (right) the William C. Rose Award at the ASBMB meeting in 2005. (Note: Prof. Rose was Prof. Coon’s mentor, so this award represents three generations of biochemists, all affiliated at some point with the University of Illinois.)

Figure 5

Immunoblotting in 1981 to 1982 (16). The left side shows an electrophoretogram (7.5% acrylamide) of samples of liver microsomes prepared from male rats treated with 2-acetylaminofluorene (AAF), trans-stilbene oxide, (SO), β-naphthoflavone (BNF), 3-methylcholanthrene (3-MC), Aroclor 1254 (AROCLOR), N,N-dimethylnitrosamine (DMN), 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), corn oil only (CONTROL), pregenenolone-16α-carbonitrile (PCN), or phenobarbital (PB) and stained using what would today be termed rabbit anti-rat P450 1A1 (then termed BNF-B₂). The panel on the right side shows the integrals of scans of the peaks (inset) detected by rabbit anti-rat P450 2B1 (then PB-B₂) in liver microsomes of male rats treated with phenobarbital to induce the enzyme. Note the linearity of the response (open and closed circles are replicates) up to 5 μg protein. In this early work, horseradish peroxidase and benzidine were used to stain the immunoreactive proteins.

Figure 6

JBC associate editors at their meeting in San Francisco in February 2010.Back (left to right): Vince Hascall, Jerry Lingrel, Jim Stull, Ken Neet, David Russell, Chuck Samuel, the author, Jim Siedow, John Exton, Bob Lehman, Bob Simoni, and George Carman. Front (left to right): Norma Allewell, Bill Smith, Linda Spermulli, Tom Vanaman, Herb Tabor, Dale Benos, Judy Bond, Joel Gottesfeld, Martha Fedor, and Xiao-Fan Wang. Unfortunately, Drs Neet, Siedow, Exton, Simoni, Tabor, and Benos are no longer with us (deceased). (Note: figure previously published in JBC—W.L. Smith Reflection, 2019, volume 294, issue 6, pp. 1779–1793.)

Figure 7

Lunch at Herb Tabor’s home on the NIH campus (September 2019).Left to right: Herb Tabor, Nancy Rodnan (then ASBMB Director of Publications), and the author.

Figure 8

Some former postdocs and students at a meeting in Fukuoka, Japan in December 2012 celebrating the 50th anniversary of the discovery of P450 (128) and honoring Prof. Tsuneo Omura (note the unfortunate deletion of the “o” in “Cytochrome”).Left to right: Raku Shinkyo, Yuki Nishimura, Hideaki Sato, Yume-Fang Ueng, Chul-Ho Yun, Shin’ichi Yoshihara, Masahiko Iwasaki, the author, Kengo Watanabe, Nobuyuki Koga, and Hiroshi Yamazaki.

Figure 9

Former graduate students, postdocs, etc. at a Dog Dinner at the 2015 ICCP450 meeting in Tokyo.Front row, left to right: Liz Gillam, Yasuo Seto, Tsutomu Shimada, Osamu Okazaki, the author, Hiroshi Yamazaki, Yukari Yamazaki, Laura Furge, Chul-Ho Yun. The Guengerich lab has the nickname of the “Dogs,” going back to the late 1980s. Reunion dinners are often held at scientific conferences and referred to as “Dog Dinners.”

Figure 10

Participants in the Guengerich lab reunion celebration at Vanderbilt (October 2015). Attendees came to Nashville from 7 countries over 4 continents. Front row: Tom Harris, Anna Guengerich (daughter), Cheryl Guengerich (wife), the author, Laura Guengerich (daughter), Tsutomu and Setsuno Shimada, and Dan Liebler.

Figure 11

A group meal (“Dog Dinner”) and celebration with some current and previous graduate students and postdocs.This wasat the 21st ICCP450 meeting (Brisbane, Queensland, Australia, 2019).

Figure 12

A picture of a tree depicting my role (FPG initials on trunk), growing out of the roots of my mentors and their mentors (101) and generating the branches and leaves of my students and postdocs, with their initials. Presented at the 2019 ICCP450 meeting in Brisbane, Australia and now hanging in the author’s office. Thanks again to Liz Gillam for the concept.

Figure 13

The author (left) and his three children (left to right: Anna, Phillip, Laura).The photo was taken (by author) after a hike to the summit of the Grace Ridge Trail in Kachemak Bay State Park, Alaska in 2020 (1000 m elevation).

Figure 14

The author working with some of his equipment. This photography was takenin New Zealand (April 2023).

Figure 15

At the ASBMB office with one of the author’s pictures (taken in Okinawa at the ICCP450 2009 meeting). The author and Nancy Rodnan.