Proving the Hormonal Nature of Nerve Growth Factor: the Work of Ralph A. Bradshaw
Year: 2008
Type: article
Abstract: Artifacts in Studies of Protein Synthesis with Radioactive Amino Acids: INVALIDITY OF ALLEGED STIMULATION OF PROTEIN SYNTHESIS BY THYROXINE IN ABSENCE OF MITOCHONDRIAJournal of Biological ChemistryVol. 249Issue 17PreviewPrevious studies have demonstrated that thyroid hormones in vitro stimulate microsomal protein synthesis in cell-free systems, but the effect was found to be indirect and dependent on the presence of mitochondria in the incubation system. The mitochondrial requirement has recently been questioned by Carter et al. ((1971) J. Biol. Chem. 246, 4973), who claimed to observe direct stimulations by thyroxine in cell-free liver systems lacking mitochondria. The system used by them contained higher Mg2+ concentrations, lower microsomal concentrations, and uniformly labeled l-[14C]valine of very high specific activity. Full-Text PDF Open AccessProperties and Specificity of Binding Sites for 125I-Nerve Growth Factor in Embryonic Heart and BrainJournal of Biological ChemistryVol. 249Issue 18PreviewThe specific binding of 125I-nerve growth factor (125I-NGF) to the low speed pellet fraction of dissociated 13-day chick embryo heart and brain is a complex, nonsaturable process exhibiting a range of affinities (heart,Ka = 2.9 x 109 to 6 x 106 liters per mole and brain, 2.5 x 109 to 8 x 106 liters per mole). The binding to both tissues at 24° is rapid (k+1 = 7 x 106m-1s-1) and reversible (k-1 = 7 x 10-4s-1) and the dissociation of bound 125I-NGF is accelerated by 8 x 10-8m native NGF (k-1 = 4 x 10-3s-1). Full-Text PDF Open Access Properties of the Specific Binding of 125I-Nerve Growth Factor to Responsive Peripheral Neurons (Frazier, W. A., Boyd, L. F., and Bradshaw, R. A. (1974) J. Biol. Chem. 249, 5513–5519) Properties and Specificity of Binding Sites for 125I-Nerve Growth Factor in Embryonic Heart and Brain (Frazier, W. A., Boyd, L. F., Pulliam, M. W., Szutowicz, A., and Bradshaw, R. A. (1974) J. Biol. Chem. 249, 5918–5923) Ralph A. Bradshaw was born in Dorchester, Massachusetts in 1941. He attended Colby College in Waterville, Maine, where he majored in chemistry. During his senior year at Colby, he took a course in biochemistry taught by Wilmon B. Chipman, a recent graduate of the University of Illinois and a new addition to the Colby faculty. Chipman was young and enthusiastic about biochemistry, and less than 2 weeks into the course Bradshaw decided that he would go to graduate school to pursue studies in biochemistry. He applied to several schools but settled on Duke University because of the prominent faculty in the biochemistry department and because Duke offered the most financial support. Almost immediately after arriving at Duke, Bradshaw decided he wanted to work for Journal of Biological Chemistry (JBC) Classic author Robert L. Hill (1Brew K. Castellino F.J. Vanaman T.C. Hill R.L. J. Biol. Chem. 1970; 245 (JBC Classics:): 4570-4582Abstract Full Text PDF PubMed Google ScholarVanaman T.C. Brew K. Hill R.L. J. Biol. Chem. 1970; 245: 4583-4590Abstract Full Text PDF PubMed Google ScholarTrayer I.P. Hill R.L. J. Biol. Chem. 1971; 246: 6666-6675(http://www.jbc.org/cgi/content/full/281/6/e6)Abstract Full Text PDF PubMed Google Scholar). Hill had only come to Duke the year before Bradshaw arrived, but he already had a sizable research group that focused on the structure-function relationships of proteins. From the biochemistry course taught by Chipman, Bradshaw knew that this was where his interests lay and he “was thrilled beyond words” (2Bradshaw R.A. How I became a biochemist.IUBMB Life. 2006; 58: 495-498Crossref PubMed Scopus (0) Google Scholar) when Hill accepted him into his group. Working with Hill, Bradshaw characterized hemoglobin from four species of lemur, defined active site residues in fumarase using chemical modifications, and used cysteine-mixed disulfide derivatives to define aspects of disulfide bond formation in lysozyme. Bradshaw received his Ph.D. in 1966 and then spent a year doing a postdoctoral fellowship with Frank Gurd at Indiana University where he sequenced two aquatic mammal myoglobins. After that he joined JBC Classic author Hans Neurath (3Schwert G.W. Neurath H. Kaufman S. Snoke J.E. J. Biol. Chem. 1948; 172 (JBC Classics:): 221-239Abstract Full Text PDF PubMed Google ScholarDavie E.W. Neurath H. J. Biol. Chem. 1955; 212: 515-530(http://www.jbc.org/cgi/content/full/280/2/e1)Abstract Full Text PDF PubMed Google Scholar) at the University of Washington where he completed the sequence of bovine carboxypeptidase A. In 1969, Bradshaw was offered a position on the faculty of the Department of Biological Chemistry at Washington University School of Medicine in St. Louis. He accepted the offer knowing that the position would provide ample opportunity for productive collaborations. Sure enough, soon after arriving in St. Louis, Bradshaw, in collaboration with Rita Levi-Montalcini, determined the sequence of mouse nerve growth factor (NGF) with Ruth Angeletti (4Angeletti R.H. Bradshaw R.A. Nerve growth factor from mouse submaxillary gland: amino acid sequence.Proc. Natl. Acad. Sci. U. S. A. 1971; 68: 2417-2420Crossref PubMed Scopus (304) Google Scholar). This initial collaboration with Levi-Montalcini, who was awarded the Nobel Prize in 1986 for her pioneering work in discovering NGF, led Bradshaw to spend more than 35 years studying growth factors, their receptors, and their signaling mechanisms. Some of his NGF receptor studies are the subject of the two JBC Classics reprinted here. Both papers are binding studies that showed that iodinated NGF bound to NGF-responsive cells (peripheral neurons) with high affinity and specificity, thus establishing that they had cell surface receptors and were hormonal in nature, an idea that directly arose from the amino acid sequence determination. In the first paper Bradshaw and his colleagues study the interaction of NGF with its receptors on the surface membranes of responsive chick embryo sympathetic and sensory neurons and report that the binding is a complex process with properties similar to the binding of insulin to fat and liver cell membranes and human lymphocytes. In the second Classic, Bradshaw and his colleagues continue their binding studies, this time using embryonic heart and brain tissues from chickens, and report that binding in these tissues is very similar to that in the sympathetic and sensory nervous systems. This latter study presaged the much wider range of responsive tissues for NGF, particularly the central nervous system, that was eventually established several years later. The results of these two papers shook up the field as, up until then, NGF and epidermal growth factor, which had been discovered in the mouse submandibular gland by JBC Classic author Stanley Cohen (5Cohen S. J. Biol. Chem. 1962; 237 (JBC Classics:): 1555-1562Abstract Full Text PDF PubMed Google ScholarSavage Jr., C.R. Inagami T. Cohen S. J. Biol. Chem. 1972; 247: 7612-7621(http://www.jbc.org/cgi/content/full/281/10/e10)Abstract Full Text PDF PubMed Google Scholar) while studying NGF, were not even considered to be related to hormones. Immediately, many other substances such as platelet-derived growth factor, the somatomedins (now insulin-like growth factors), the transforming growth factors, and the fibroblast growth factors shot to center stage, and the growth factor field, as an expansion of endocrinology, was born with a vengeance. Bradshaw and colleagues eventually characterized and isolated the NGF receptor, TrkA (6Costrini N.V. Kogan M. Kukreja K. Bradshaw R.A. Physical properties of the detergent-extracted nerve growth factor receptor of sympathetic ganglia.J. Biol. Chem. 1979; 254: 11242-11246Abstract Full Text PDF PubMed Google Scholar, 7Massague J. Guillette B.J. Czech M.P. Morgan C.J. Bradshaw R.A. Identification of a nerve growth factor receptor protein in sympathetic ganglia membranes by affinity labeling.J. Biol. Chem. 1981; 256: 9419-9424Abstract Full Text PDF PubMed Google Scholar, 8Kouchalakos R.N. Bradshaw R.A. Nerve growth factor receptor from rabbit sympathetic ganglia membranes: relationship between subforms.J. Biol. Chem. 1986; 261: 16054-16059Abstract Full Text PDF PubMed Google Scholar), substantially before it was cloned. In 1982, Bradshaw moved to California where he became Professor and Chair of the Department of Biological Chemistry at the University of California, Irvine. He is currently Professor Emeritus in the Department of Physiology & Biophysics at the University of California, Irvine as well as Professor in the Department of Pharmaceutical Chemistry and Deputy Director of the Mass Spectrometry Facility at the University of California, San Francisco. Bradshaw has been very active in the American Society for Biochemistry and Molecular Biology, serving on the Committee on Equal Opportunities for Minority Groups, the Nominating Committee, and the Membership Committee and also acting as Society Treasurer and Councilor. He is currently Deputy Chair of the ASBMB Public Affairs Advisory Committee and Society Historian. Bradshaw was also the President of the Federation of American Societies for Experimental Biology, Treasurer of the Keystone Symposia, and the founding President of the Protein Society. He has served on many editorial boards and was an Associate Editor of the Journal of Biological Chemistry. He is currently Co-editor of Molecular and Cellular Proteomics. In recognition of his contributions to science, Bradshaw has received many awards and honors including the Passano Foundation Young Scientist Award (1976), the American Chemical Society St. Louis Section Award (1979), the Italian National Research Council Gold Medal (1987), and the Australia Society of Medical Research Gold Medal (1999). 1Biographical information on Ralph Bradshaw was taken from Ref. 2Bradshaw R.A. How I became a biochemist.IUBMB Life. 2006; 58: 495-498Crossref PubMed Scopus (0) Google Scholar. 1Biographical information on Ralph Bradshaw was taken from Ref. 2Bradshaw R.A. How I became a biochemist.IUBMB Life. 2006; 58: 495-498Crossref PubMed Scopus (0) Google Scholar., 2We would like to thank Ralph Bradshaw for providing background information for this introduction. 2We would like to thank Ralph Bradshaw for providing background information for this introduction. The first author of both the Bradshaw Classics, William A. Frazier, has also gone on to have a very productive career in science. At the time the papers were published, Frazier was a predoctoral student in Bradshaw's laboratory. A few years later, he became an Assistant Professor in the Department of Biological Chemistry at Washington University School of Medicine. He remains there today and is currently a Professor of Cell Biology and Physiology and Professor of Biomedical Engineering. His research focuses on the function, structure, and mechanism of thrombospondin-1 (TSP1) and its receptors, CD36 and CD47.
Source: Journal of Biological Chemistry
Language: English
Subfield: Cellular and Molecular Neuroscience
Field: Neuroscience
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $3,430
OpenAlex ID: W1581495852