Biophysics of Signal Transduction


Signal transduction networks are key processes utilized by organisms to sense their environment and to regulate essentially all life processes such as their metabolism, motility, reproduction, etc. Physically, signal transduction proceeds via local initial changes upon stimulation – e.g. light absorption or ligand binding followed by elementary events such as electron transfer, hydrogen-bond rearrangement, isomerization. In turn, the Van der Waals complementarities that thus far defined the protein’s conformation are disturbed, leading to larger-scale movements of the protein and, ultimately in changes of affinity of interacting signaling proteins. Over the last years I have developed a research program at the VU laser center to unravel the molecular mechanisms of light-driven signal transduction. The goal of the program is to understand the dynamic-structural basis of signal transduction at the molecular level, from initiation by a flash of light to propagation of information-encoding structural changes in the signaling proteins, to organismal output. To this end, advanced time-resolved spectroscopic techniques such as ultrafast transient absorption, ultrafast IR and Raman, time-resolved FTIR and multi-pulse spectroscopy are employed.


Signal transduction proteins and networks have a strictly modular architecture with conserved input and output domains. Light-driven signaling proteins and networks often are similar to those that have quite different input signals (such as oxygen, temperature, food sources, metabolites etc.) and are eminently suited for biophysical studies since they can be activated by a short laser flash. The model systems of the program are mainly bacterial in origin and chosen in such a way that knowledge with a high degree of general significance is obtained. By studying the intact photo-activated signal transduction proteins at increasing levels of complexity, to culminate in light-driven signal transduction studies in whole cells, the research is firmly placed in a Systems Biology context. Moreover, the light-sensitive input domains are photochromic and photoreversible (i.e., they can be turned ‘on’ and ‘off’ by laser flashes of different color) which offers unprecedented control over signal transduction pathways by means of multi-pulse control spectroscopies.


The program carries a considerable significance for the society at large. The bacterial model signaling proteins and networks of the program have many homologues in human pathogens. The latter form a prime target for the development of new generations of antibiotics, which are dearly needed in the face of ever increasing resistance to existing ones. Likewise, several forms of cancer are caused by disturbances in human signaling pathways analogous to those under investigation in the program, and may be cured by intervention at the molecular level.

Presently the program is supported by grants from NWO-ALW: a VIDI grant to myself (600 kEur), the Molecule to Cell program (500 kEur, with R. van Grondelle and K.J. Hellingwerf) and an investment grant to construct an advanced multi-pulse spectrometer (672 kEur, with R. van Grondelle). Close collaborations exist with prof. Klaas Hellingwerf at SILS/UvA, with prof. Sean Crosson at the University of Chicago, prof. Peter Hegemann at the Humboldt University, Berlin and professor Keith Moffat at the University of Chicago.

 

Key publications

 
J.T.M. Kennis and S. Crosson
A bacterial pathogen sees the light
Science 317, 2007, p. 1041-1042 (Perspective)

M.T.A. Alexandre, J.C. Arents, R. van Grondelle, K.J. Hellingwerf, J.T.M. Kennis
A base-catalyzed mechanism for dark state recovery in the Avena sativa phototropin-1 LOV2 domain
Biochemistry
46, 2007, p. 3129-3137

M. Gauden, J.S Grinstead, W. Laan, I.H.M. van Stokkum, M. Avila-Perez, K.C. Toh, R. Boelens, R. Kaptein, R. van Grondelle, K.J. Hellingwerf, J.T.M. Kennis
On the role of aromatic side chains in the photoactivation of BLUF domains
Biochemistry 46, 2007, p. 7405-7415

M. Gauden, I.H.M. van Stokkum, J.M. Key, D. Ch. Luhrs, R. van Grondelle, P. Hegemann, J.T.M. Kennis, Hydrogen bond switching via a radical pair mechanism in a flavin-binding photoreceptor Proc. Natl. Acad. Sci. USA 103, 2006, p. 10895-10900

 M. Gauden, S. Yeremenko, W. Laan, I.H.M. van Stokkum, J.A. Ihalainen, R. van Grondelle, K.J. Hellingwerf, J.T.M. Kennis, Photocycle of the flavin-binding photoreceptor AppA, a bacterial transcriptional anti-repressor of photosynthesis genes Biochemistry 44, 2005, p. 3653-3662

 J.T.M. Kennis, I.H.M. van Stokkum, S. Crosson, M. Gauden, K. Moffat, R. van Grondelle, The LOV2 domain of phototropin: A reversible photochromic switch
J. Am. Chem. Soc. 126, 2004, p. 4512-4513

 J.T.M. Kennis, S. Crosson, M. Gauden, I.H.M. van Stokkum, K. Moffat, R. van Grondelle, Primary reactions of the LOV2 domain of phototropin, a plant blue-light photoreceptor Biochemistry 42, 2003, p. 3385-3392