... understanding life in molecular detail

Prof Peter Henderson

Membrane transport, antibiotic resistance


Structures and mechanisms of membrane transport proteins, antibiotic resistance, drug development, developing applications of physical  methods to elucidate structure-activity relationships (SAR) of membrane proteins.

Current major projects include:
  • SAR of the Mhp1 transport protein.
  • Efflux proteins confering drug resistance in pathogenic bacteria
  • Developing NMR and MS for elucidating SAR of transport proteins
  • Application of NMR for elucidating SAR of membrane proteins.

The laboratory is concerned with elucidating the structure-activity relationships of a wide range of membrane transport proteins found in bacteria. Included are uptake proteins for sugars and amino acids and efflux proteins for antibiotics from 'model' organisms like Escherichia coli, Bacillus subtilis, Streptomyces coelicolor and Corynebacterium glutamicum, and an increasing number of important pathogens such as Helicobacter pylori, Brucella melitensis, Staphylococcus aureus and others.

Our strategy is to transfer genes encoding transport proteins to suitable vectors for amplification of expression in E. coli. A number of the proteins are homologues to those found in man, so the bacteria provide an highly convenient model for elucidating the molecular mechanisms of mammalian transporters.

Proteins purified in 2-20 mg quantities are used for both 2D and 3D crystallisation trails and a variety of physical measurements, fluorimetry, MS, NMR, CD and FTIR and EPR spectroscopy. Mutants and/or chimaeric proteins are also examined to locate substrate/inhibitor binding sites, elucidate the translocation process, facilitate structural studies, etc.

A major goal is to determine the three-dimensional structure(s) of the proteins in the membrane using electron crystallography, X-ray crystallography and NMR. The proteins are being produced with cysteine, tryptophan and other residues engineered to facilitate these and other approaches. Working with Dr Alex Cameron at Warwick and the Membrane Protein Laboratory at the Diamond Synchrotron, the structure of an sodium-hydantoin transport protein was determined in three conformations, open-out, occluded-with-substrate, and open-inwards (Figure 1). This enabled the structural basis of the alternating access mechanism of transport to be visualized for the first time. In addition genetical manipulation is used to determine which regions of each protein are involved in substrate recognition, cation recognition, inhibitor binding, translocation, Km, Vmax, Ki, etc.

Individual amino acid residues are changed as informed by our databases of aligned transporter sequences.

Figure 1. Outward- and inward- facing cavities in Mhp1.  A slice through the Mhp1 structures, viewed parallel to the membrane is shown.  (A) Mhp1 outward-open structure (substrate-free).  (B) Mhp1 substrate-occluded with benzylhydantoin bound.  (C) Mhp1 the open-inward cavity is shown.  The benzylhydantoin is shown in cyan.  Diagram taken from Shimamura et al. (2010).

Detailed research programme                  Close ▲
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Professor of Biochemistry and Molecular Biology 1992-present
BSc (Bristol) PhD (Bristol)
1968-1971 Fulbright-Hays Scholarship for research at the University of Wisconsin, USA; 1982-1983 Visiting Professorship at the Jichi Medical School, Japan, sponsored by the Royal Society and the Japan

Postdoc, Madison, Wisconsin 1968-1971
Lecturer, University of Leicester 1973-1975
Lecturer, University of Cambridge 1975-1990
Reader, University of Cambridge 1990-1992

Astbury 6.108b
School of Molecular and Cellular Biology
0113 343 3175
p.j.f.henderson@leeds.ac.uk

Selected Publications

  1. Pikyee, M., Patching, S.G., Ivanova, E., Baldwin, J.M., Sharples, D., Baldwin, S.A. and Henderson, P.J.F. (2016) “The allantoin transport protein, PucI, from Bacillus subtilis: evolutionary relationships, amplified expression, activity and specificity”. Microbiology 162,  823-836.

  2. Hassan, K.A., Liu, Q., Henderson, P.J.F. and Paulsen, I.T. (2015) “Homologs of the Acinetobacter baumannii AceI transporter represent a new family of bacterial multidrug efflux systems”. mBio 6 (1) e01982-14, 1-5

  3. Simmons, K.J., Jackson, S.M., Brueckner, F., Patching, S.G., Beckstein, O., Ivanova, E., Geng, T., Weyand, S., Drew, D., Lanigan, J., Sharples, D., Sansom, M.S.P., Iwata, S., Fishwick, C.W.G., Johnson, A.P., Cameron, A.D. and Henderson, P.J.F. (2014) “Molecular mechanism of ligand recognition by membrane transport protein, Mhp1”. EMBO J 33, 1831-1944

  4. Hassan, K.A., Jackson, S.M., Penesyan, A., Patching, S.G., Tetu, S.G., Eijkelkamp, B.A., Brown, M.H., Henderson, P.J.F. and Paulsen, I.T. (2013) Transcriptomic and biochemical analyses identify a novel family of chlorhexidine efflux proteins”. Proc Natl. Acad. Sci. USA. 110 (50), 20254-20259