Association of a purine-analogue-sensitive protein kinase activity with p75 nerve growth factor receptors

C Volonte, AH Ross and LA Greene

Department of Pathology, College of Physicians and Surgeons, Columbia University, New York, New York 10032.

Purine analogues are protein kinase inhibitors, and they block with varying potency and specificity certain of the biological actions of nerve growth factor (NGF). The analogue 6-thioguanine (6-TG) has been shown to inhibit with high specificity protein kinase N (PKN), a serine/threonine protein kinase activated by NGF in several cellular systems. In the present work, immunoprecipitates of p75 NGF receptors from PC12 cells (+/-NGF treatment) were assayed for protein kinase activity using the substrate myelin basic protein under phosphorylating conditions optimal for PKN and in the presence or absence of purine analogues. An NGF-inducible activity was detected, and approximately 80% was inhibited by purine analogues. This activity was maximally stimulated by NGF within 5-10 min, partially decreased by 60 min, and returned to basal levels after 15 h of NGF treatment. The analogue 6-TG inhibited the NGF-inducible p75-associated kinase activity with an IC50 in the range of 15-35 microM. In mutant PC12 nnr-5 cells that lack the Trk NGF receptor, the purine-analogue-sensitive p75-associated kinase activity was not inducible by NFG. In normal PC12 cells, cyclic AMP analogues and epidermal growth factor failed to induce the same activity. Application of either 2-aminopurine or 6-TG to intact cells only slightly inhibit the NGF-dependent induction of the purine- analogue-inhibited p75-associated kinase activity. This activity shares many similarities but also displays some significant differences with cytosolic PKN. Our findings therefore indicate the association of a purine-analogue-sensitive protein kinase with p75 NGF receptors.

Volume 4, Issue 1, pp. 71-78, 01/01/1993
Copyright © 1993 by The American Society for Cell Biology

This article has been cited by other articles:

				K D S. A. Wansa and G. E O Muscat TRAP220 is modulated by the antineoplastic agent 6-Mercaptopurine, and mediates the activation of the NR4A subgroup of nuclear receptors J. Mol. Endocrinol., June 1, 2005; 34(3): 835 - 848. [Abstract] [Full Text] [PDF]

				Y. Denkins, J. Reiland, M. Roy, N. D. Sinnappah-Kang, J. Galjour, B. P. Murry, J. Blust, R. Aucoin, and D. Marchetti Brain metastases in melanoma: Roles of neurotrophins Neuro-oncol, April 1, 2004; 6(2): 154 - 165. [Abstract] [PDF]

				P. Ordentlich, Y. Yan, S. Zhou, and R. A. Heyman Identification of the Antineoplastic Agent 6-Mercaptopurine as an Activator of the Orphan Nuclear Hormone Receptor Nurr1 J. Biol. Chem., June 27, 2003; 278(27): 24791 - 24799. [Abstract] [Full Text] [PDF]

				S. Lei, W. F. Dryden, and P. A. Smith Involvement of Ras/MAP Kinase in the Regulation of Ca2+ Channels in Adult Bullfrog Sympathetic Neurons by Nerve Growth Factor J Neurophysiol, September 1, 1998; 80(3): 1352 - 1361. [Abstract] [Full Text] [PDF]

				D. E. Wolf, C. A. McKinnon, M.-C. Daou, R. M. Stephens, D. R. Kaplan, and A. H. Ross Interaction with TrkA Immobilizes gp75 in the High Affinity Nerve Growth Factor Receptor Complex J. Biol. Chem., February 3, 1995; 270(5): 2133 - 2138. [Abstract] [Full Text] [PDF]

				K. Lee, A. Davies, and R Jaenisch p75-deficient embryonic dorsal root sensory and neonatal sympathetic neurons display a decreased sensitivity to NGF Development, January 4, 1994; 120(4): 1027 - 1033. [Abstract] [PDF]