Professor John Bowie

Research Interests

1. Biologically-active host-defence peptides

Recent work in this area has involved defence peptides isolated from a wallaby, a number of Australian frogs and a spider from South America. For example:

(a) The lactating female of the Tammar wallaby (Macropus eugenii) produces a potent peptide (eugenin) which enhances killer T cell formation in her pouch protecting the young from bacterial flora and other predators [1]. Similar activity is found in disulfide containing peptides from the skin secretions of a number of froglets of the genus Crinia, e.g. riparin 1 [2].

Eugenin       pEQDY(SO₃H)VFMHPF-NH₂
Riparin 1      RLCIPVIFPC-OH
Caerin 1.8    GLFKVLGSVAKHLLPHVVPVIAEKL-NH₂

(b) As well as antibiotics and neuropeptides, Australian frogs contain peptides in their skin glands, which, when secreted onto their skin inhibit the production of the chemical messenger nitric oxide by the nitric oxide synthase isoforms. Ingestion of these peptides will either incapacitate or kill a predator. The peptides act by complexing with the regulatory peptide (molecular switch) Ca²⁺ calmodulin which is then unable to attach to the calmodulin docking domain of the appropriate nitric oxide synthase [3]. The helix-hinge-helix peptide caerin 1.8, is one of the most potent of this class of peptide. This work has been reviewed [2,4].

We are currently investigating, together with Prof. John Carver and Dr Tara Pukala, whether any of the peptides that we have isolated from the skin glands of frogs and toads, inhibit the aggregation of those peptides which are responsible for the formation of the fibrils responsible for Alzheimers and Parkinson diseases.

(c) The venom of the South American spider Cupiennius salei, contains four 35 residue cupiennin peptides. These peptides, e.g. cupiennin 1a, are known antimicrobial agents, but it has now shown that these peptides are more potent in deactivating NOS than the amphibian peptides described above, and that they also operate by complexing with calmodulin. This assists with paralysis of the prey following envenomation [5,6].

Cupiennin 1a   GFGALFKFLAKKVAKTVAKQAAKQGAKYVVNKQME-NH₂

2. Ion chemistry (mass spectrometry)

A major research interest over the years has been the study of negative ion mass spectrometry. Our recent work in this area is currently involved in the use of negative ions to (i) to form neutrals of stellar significance [7,8], (ii) develop the use of negative ion mass spectrometry as a viable analytical method for the sequence determination of peptides and proteins [9,10], and (iii) to use ion-molecule chemistry of transient anions to study the structure and reactivity of these anions in the gas phase.

One of the most important questions in science is how did life originate on our planet. Hoyle proposed [11] that the primary chemicals of life originate in space, i.e. by reactions in interstellar dust clouds, circumstellar envelopes or interstellar ice. We strip an electron from negative ions in the mass spectrometer to produce a reactive neutral of known bond connectivity. We study the structure and reactivity of this neutral in the mass spectrometer in order to assess the likelihood that it is long-lived enough to be able to undergo reaction to form an amino acid, a sugar or a nucleotide base. As an example, we have shown that electron stripping of ^-CH₂CN produces the stable radical ^.CH₂CN which may then produce the glycine precursor NH₂CH₂CN which can then be hydrolysed in interstellar ice (around comets, planets etc) to yield the amino acid glycine [12]. We are currently considering ways by which other amino acids can be formed in stellar regions.

^.CH₂CN + NH_2^. → NH₂CH₂CN → NH₂CH₂CO₂H (glycine)

The prototypical two carbon sugar glycolaldehyde (HOCH2CHO) has been detected towards the galactic centre source Sagittarius B2(N) by means of rotational transitions [13].  We are studying, by experiment and theory in concert, the possible interstellar synthetic pathways to form glycolaldehyde and higher sugars.

We are also interested in cumulenes that have been detected in interstellar molecular clouds and are currently studying linear CCCX and their planar rhombic isomers. These are formed by the charge stripping of negative ions in a mass spectrometer, e.g. CCCC [14] and CCCN [15]. This experimental work is complemented by theoretical studies (computational chemistry) in order to probe the structure and reactivity of these transient cumulenes.

Finally, we are currently using ion molecule chemistry in a modified ion-trap mass spectrometer to investigate the structures of transient carbanions in the gas phase; e.g. the reaction between a carbanion and carbon disulfide to form an adduct whose subsequent fragmentations provide information concerning the structure of the original carbanion.  This work is also complemented by theoretical studies.

References

1	R.V.Baudinette, P.Boontheung, I.F.Musgrave, P.A.Wabnitz, V.M.Maselli, J.Skinner, P.F.Alewood, C.S.Brinkworth and J.H.Bowie, FEBS J., 2005, 272, 433-443.
2	T.L.Pukala, J.H.Bowie, V.M.Maselli, I.F.Musgrave and M.J.Tyler, Nat. Prod. Rep., 2006, 23, 368-393.
3	J.Doyle, C.S.Brinkworth, K.L.Wegener, J.A.Carver, L.E.Llewellyn, I.N.Olver, J.H.Bowie, P.A.Wabnitz and M.J.Tyler, Eur. J. Biochem., 2003, 270, 1141-1153.
4	M.A.Apponyi, T.L.Pukala, C.S.Brinkworth, V.M.Maselli, J.H.Bowie, M.J.Tyler, G.W.Booker, J.C.Wallace, J.A.Carver, F.Separovic, J.R.Doyle and L.E.Llewellyn. Peptides, 2004, 25, 1035-1054.
5	T.L.Pukala, J.R.Doyle, L.E.Llewellyn, L.Kuhn-Nerwig, F.Separovic and J.H.Bowie. FEBS J., 2007, 274, 1778-1784..
6	T.L.Pukala, M.P.Boland, J.D.Gehman, L.Kuhn-Nerwig, F.Separovic and J.H.Bowie. Biochemistry, 2007, 46, 3576-3585.
7	S.J.Blanksby and J.H.Bowie, Mass Spectrom. Rev. 1999, 18, 131-151.
8	S.Dua, S.J.Blanksby, S.Peppe, A.M.McAnoy and J.H.Bowie, Curr. Org. Chem., 2003, 7, 1545-1564.
9	J.H.Bowie, C.S.Brinkworth and S.Dua, Mass Spectrom. Rev.,2002, 21, 87-107.
10	D.Bilusich and J.H.Bowie, Rapid Commun. Mass Spectrom., 2007, 21, 619-628.
11	E.M.Burbidge, G.R. Burbidge, W.A. Fowler and F. Hoyle. Revs. Mod. Physics 1957, 29, 547-650.
12 13	H.J.Andreazza, M.Fitzgerald and J.H.Bowie, Org. Biomol. Chem., 2006, 4, 2466-2472. J.M.Hollis, F.J.Lovas and P.R.Jewell, Astrophys. J., 2000, 540, L107.
14	S.J.Blanksby, D.Schröder, S.Dua, J.H.Bowie and H.Schwarz, J. Am. Chem. Soc., 2000, 122, 7105-7113.
15	M.J.Maclean, M.Fitzgerald and J.H.Bowie. J. Phys. Chem. A, 2007, 111, 12932-12937.

Publications

Research awards

  E.H.Rennie Medal, 1967 (Royal Australian Chemical Institute).

  H.G.Smith Medal, 1973 (Royal Australian Chemical Institute).

  Morrison Medal, 1997 (Australian and New Zealand Society for Mass Spectrometry)

  A.J.Birch Medal, 2001 (Organic Division, Royal Australian Chemical Institute)

  Sir J.J.Verco Medal, 2003 (Royal Society of South Australia)

  J.J.Thomson Gold Medal, 2006 (International Council for Mass Spectrometry).

 

Publications 2005 -

S.J.Blanksby and J.H.Bowie Carbanions: formation, structure and reactivity. In Encyclopaedia of Mass Spectrometry. Ed. N.M.M.Nibbering, Vol 5, pp. 261-269. Pergamon Press, Oxford, 2005

J.H.Bowie and C.S.Brinkworth Negative ion mass spectra of underivatised peptides: an aid to sequence determination. In Encyclopaedia of Mass Spectrometry, Ed. N.M.M.Nibbering, Vol. 5, pp. 423-433. Pergamon Press, Oxford, 2005.

J.H.Bowie Carbanions and enolate anions. Collision induced fragmentations: an aid to structure determination. In Encyclopaedia of Mass Spectrometry. Ed., N.M.M.Nibbering, Vol. 5, pp.443-449. Pergamon Press, Oxford, 2005.

J.H.Bowie Reactions of anions with carbonyl centers. C-C bond forming reactions. In Encyclopaedia of Mass Spectrometry. Ed. N.M.M.Nibbering, Vol. 5, pp. 470-474. Pergamon Press, Oxford, 2005.

R.V.Baudinette, P.Boontheung, I.F.Musgrave, P.A.Wabnitz, V.M.Maselli, P.F.Alewood , C.S.Brinkworth and J.H.Bowie Eugenin: an immunomodulator used to protect young in the pouch of the Tammar Wallaby Macropus eugenii. FEBS Journal, 2005, 272, 433-443.

M.Fitzgerald, A.M.McAnoy, J.H.Bowie D.Schröder and H.Schwarz, Neutral cumulene oxide CCCCO is accessible by oxidation of [CCCCO]^-. In the gas phase. Org. Biomol. Chem., 2005, 3, 901-910.

M.Fitzgerald, S.Dua and J.H.Bowie One-electron oxidation of [CCOCC]^-. in the gas phase gives stable and decomposing forms of CCCCO. Org. Biomol.Chem., 2005, 3, 2646-2651,

E.Ambroglio, F.Separovic, J.H.Bowie, G.D.Fidelio and L.A.Bagatoli Direct visualization of membrane leakage by antibiotic peptides maculatin 1.1, citropin 1.1 and aurein 1.1. Biophys. J., 2005, 89, 1874-1881.

S.E.Van Compernolle, R.J.Taylor, K.Oswald-Richter, A.Posdoc, B.E.Youree, J.H.Bowie, M.J.Tyler, J.M.Conlon, D.Wade, T.Dermody, C.Aiken, L.A.Rollins-Smith and D.Unutmaz. Amphibian antimicrobial skin peptides that potently inhibit HIV infection and dendridic cell mediated transfer of virus to T cells. J. Virol., 2005, 79, 12088-12094.

C.S.Brinkworth, J.H.Bowie, D.Bilusich and M.J.Tyler The rothein peptides from the skin secretion of Roth's tree frog. Sequence determination using positive and negative ion electrospray mass spectrometry. Rapid Commun. Mass Spectrom., 2005, 19, 2716-2724.

V.V.Lobodin, A.T.Lebedev, Y.Y.Morzherin, T.Blumenthal, D.Bilusich, J.H.Bowie and V.A.Ovacharenko The gas phase cyclisation of deprotonated N-aryl-2-diazo-2-cyanoacetamides. ARKIVOC 2005 (iv) 189-198.

D.Bilusich, V.M.Maselli, C.S.Brinkworth, T.Samguina, A.T.Lebedev and J.H.Bowie Direct identification of disulfide links in peptides using negative ion electrospray mass spectra of underivatised peptides. A joint experimental and theoretical study. Rapid Commun. Mass Spectrom., 2005, 19, 3063-3074.

M.Fitzgerald, J.H.Bowie, D.Schröder and H.Schwarz The formation of neutrals CCCO₂H and HCCCO₂ from anionic precursors in the gas phase: a joint experimental and theoretical study. Rapid Commun. Mass Spectrom., 2005, 19, 3705-3712.

S.Dua, M.J.Maclean, M.Fitzgerald, A.M.McAnoy and J.H.Bowie Is the hypothionite anion (OSCN)^- the major product in the peroxidase catalysed oxidation of the thiocyanate anion (SCN)^-? A joint experimental and theoretical study. J. Phys. Chem. A., 2006, 110, 4930-4936.

M.Fitzgerald and J.H.Bowie One electron oxidation of HCCOCC^- in the gas phase gives stable HCCOCC and decomposing HCCCCO. Rapid Commun. Mass Spectrom., 2006, 20, 577-582.

P.Boontheung, C.Gao, C.S.Brinkworth, J.H.Bowie and M.J.Tyler Is the Australian Striped Frog Litoria alboguttata, a member of Litoria or Cyclorana? A profiling study using skin peptides. Trans. Roy. Soc. South Aust., 2006, 130, 1-9.

M.J.Maclean, C.S.Brinkworth, D.Bilusich, J.H.Bowie and M.J.Tyler New Caerin antimicrobial peptides from the skin secretion of the Dainty Green Tree Frog Litoria gracilenta. Sequence determination using positive and negative ion electrospray mass spectrometry. Toxicon, 2006, 47, 664-675.

V.M.Maselli, D.Bilusich, J.H.Bowie and M.J.Tyler Host-defence peptides of the Australian Streambank Froglet Crinia riparia: isolation and sequence determination by positive and negative ion electrospray mass spectrometry. Rapid Commun. Mass Spectrom., 2006, 20, 797-803.

H.J.Andreazza, M.Fitzgerald and J.H.Bowie The formation of the radicals CH₂CN, CH₃CHCN and CH₂CH₂CN from the anions ^-CH₂CN, CH₃^-CHCN and ^-CH₂CH₂CN in the gas phase. A joint experimental and theoretical study. Org. Biomol. Chem., 2006, 4, 2466-2472.

D.Bilusich and J.H.Bowie Determination of disulfide functionality in underivatised peptides using negative ion mass spectrometry: an aid to structure determination. Current Analytical Chemistry, 2006, 2, 341-353.

J.H.Bowie and M.J.Tyler Bioactive peptides from Australian amphibians: caerulein and other neuropeptides. In A.J.Kastin, ed., The Handbook of Peptides, Academic Press, Amsterdam,  2006, Chapter 43, pp. 283-289.

T.L.Pukala, V.M.Maselli, J.H.Bowie, I.F.Musgrave and M.J.Tyler Host-defence peptides from the glandular secretions of amphibians: structure and activity. Natural Product Reports, Chemical Society (London), 2006, 23, 368-393

T.L.Pukala, T.Bertozzo, S.C.Donnellan, J.H.Bowie, K. H. Surinya-Johnson, Y.Liu, R.J.Jackway, J.R.Doyle, L.E.Llewellyn and M.J.Tyler Host defence peptide profiles of the skin secretions of interspecific hybrid tree frogs and their parents, female Litoria splendida and male Litoria caerulea. FEBS Journal 2006, 273, 3511-3519.

D.Bilusich and J.H.Bowie Direct identification of intermolecular disulfide functionality in underivatised peptides using negative ion electrospray mass spectrometry: a joint experimental and theoretical study. Rapid Commun. Mass Spectrom., 2007, 21, 619-628

T.L.Pukala, M.P.Boland, J.D.Genman, L.Kuhn-Nerwig, F.Separovic and J.H.Bowie Solution structure and interaction of cupiennin 1a, a spider venom peptide, with phospholipid bilayers. Biochemistry, 2007, 46, 3576-3585.

T.L.Pukala, J.R.Doyle, L.E.Llewellyn, L.Kuhn-Nerwig, F.Separovic and J.H.Bowie Cupiennin 1a, an antimicrobial peptide from the venom of the neotropical wandering spider Cupiennius salei, also inhibits the formation of nitric oxide by neuronal nitric oxide synthase. FEBS Journal, 2007, 274, 1778-1784.

K.Artemenko, T.Samguina, V.V.Lobodin, A.T.Lebedev, D.Bilusich and J.H.Bowie Host defence peptides from the skin secretion of the European Marsh Frog Rana ridibunda. Mass Spektrometria (Russian Society for Mass Spectrometry), 2007, 79-89.

M.J.Maclean, M.Fitzgerald and J.H.Bowie The interstellar molecule CCCN can be formed by charge stripping of [CCCN]^- in the gas phase, and undergoes carbon scrambling when energized. J. Phys. Chem.A, 2007, 111, 12932-12937.

D.C.Graham, C.Mitchell, M.I.Bruce, G.F.Metha, J.H.Bowie and M.A.Buntine Production of acrylic acid through nickel mediated coupling of ethylene and carbon dioxide: a DFT study. Organometallics, 2007, 26, 6784-6792.

J.H.Gelman, F.Luk, K.Hall. T-H.Lee, M.P.Boland. T.L.Pukala, J.H.Bowie, M-I.Aguilar and F.Separovic Effect of antimicrobial peptides from Australian tree frogs on anionic phospholipid membranes. Biochemistry, 2008, 47, 8537-8363.

A.W.Perriman, M.A.Apponyi, M.A,Buntine, R.J.Jackway, J.W.White, M.W.Rutland and J.H.Bowie Surface movement in water of splendipherin, the aquatic male sex pheromone of the tree frog Litoria splendida. FEBS Journal, 2008, 275, 3362-3374.

D.C.Graham, M.I.Bruce, G.F.Metha, J.H.Bowie and M.A.Buntine. Regioselective control of Nickel Mediated Coupling of acetylene and Carbon Dioxide - A DFT Study. J. Organometallic Chem., 2008, 693, 2703-2710.

R.J.Jackway T.L.Pukala,, V.M.Maselli, I.F.Musgrave, J.H.Bowie, Y.Liu, K.H.Surinya-Johnson, Stephen C.Donnellan, J.R.Doyle, M.J.Tyler and L.E.Llewellyn Disulfide-containing peptides from the glandular skin secretions of froglets of the genus Crinia: structure, activity and evolutionary trends. Regul. Pept., 2008, 151, 80-87.

T.L.Pukala, T.Urathamakul, S.J.Watt, J.L.Beck, R.J.Jackway and J.H.Bowie Binding studies of nNOS active amphibian peptides with Ca²⁺ calmodulin, using negative ion electrospray mass spectrometry. Rapid Commun. Mass Spectrom., 2008, 22, 3501-3507.

R.J.Jackway, J.H.Bowie, D.Bilusich, I.F.Musgrave, K.H.Surinya-Johnson M.J.Tyler and P.C.H.Eichinger The fallaxidin peptides from the skin secretion of the Eastern Dwarf Tree Frog Litoria fallax. Sequence determination by positive and negative ion electrospray mass spectrometry: antimicrobial activity and cDNA cloning of the fallaxidins. Rapid Commun. Mass Spectrom., 2008, 22, 3207-3216.

H.J.Andreazza, M.Fitzgerald, D.Bilusich, , R.Hoffmann, P.Hoffmann, P.C.H.Eichinger and J.H.Bowie Characteristic negative ion fragmentations of deprotonated peptides containing post-translational modifications: mono-phosphorylated Ser, Thr and Tyr. A joint experimental and theoretical study. Rapid Commun. Mass Spectrom., 2008, 22, 3305-3312.

M.J.Maclean, P.C.H.Eichinger, T.Wang, M.Fitzgerald and J.H.Bowie A theoretical study of the cyclisation processes of energised CCCSi and CCCP. J. Phys. Chem. A, 2008, 112, 12714-12720.

M.Fitzgerald, D.Bilusich, S.Dua, P.C.H.Eichinger, S.Peppe and J.H.Bowie Anions [N(CH₂)₃]^- and [ON(CH₂)₂]^- are stable in the gas phase, but can they be charge stripped to form the radicals N(CH₂)₃ and ON(CH₂)₂? A joint experimental and theoretical study. Eur. J. Mass Spectrom., 2009, 15, 91-104.   Dedicated to Peter J. Derrick to honour his contributions to chemistry.

 J.H.Bowie and D.Bilusich Fragmentation of (M-H)^- anions of underivatised peptides. Part 2. Characteristic cleavages of Ser and Cys, and disulfides and other post translational modifications, together with some unusual internal rearrangements. Mass Spectrom. Rev., 2009, 28, 20-34.

J.R.Doyle, L.E.Llewellyn, T.L.Pukala, R.J.Jackway and J.H.Bowie Anuran host-defence peptides which inhibit the synthesis of nitric oxide by neuronal nitric oxide synthase. In Bioactive Peptides, J.Howl and S.Jones eds., Taylor and Francis Publishing Group, U.K. 2009, Chapter 14, pp 315-332.

J.H.Bowie, R.J.Jackway, J.A.Carver, F.Separovic and M.J.Tyler Host defence peptides from the secretion of the skin glands of frogs: membrane active peptides from the genera Litoria, Uperoleia and Crinia. In Bioactive Peptides, J.Howl and S.Jones eds., Taylor and Francis Publishing Group, UK, 2009, Chapter 15, pp 333-355.

B.Gao, P.J.Sherman, L.Luo, J.H.Bowie and S.Zhu.  Structural and functional characterisation of two genetically related meucin peptides highlights evolutionary divergence and convergence in antimicrobial peptides.  FASEB Journal, 2009, 23, 1230-1245.

R.J.Jackway, V.M.Maselli, I.F.Musgrave, M.J.Maclean and J.H.Bowie.  Unusual skin peptides from anurans of the Litoria rubella Group:  sequence determination using electrospray mass spectrometry: opioid activity of two major peptides. Rapid Commun. Mass Spectrom., 2009, 23, 1189-1195.

T.Wang, D.Bilusich, H.J.Andreazza and J.H.Bowie.  Negative ion fragmentations of deprotonated peptides containing post-translational modifications.  An unusual cyclisation/rearrangement involving phosphotyrosine: a joint experimental and theoretical study.  Rapid Commun. Mass Spectrom., 2009, 23, 1669-1677.

H.J.Andreazza, T.Wang, D.Bilusich, P.Hoffmann and J.H.Bowie.  Negative ion fragmentations of deprotonated peptides containing post-translational modifications: diphosphorylated peptides containing Ser, Thr and Tyr.  A phosphate/phosphate cyclisation reaction.  A joint experimental and theoretical study.  Rapid Commun. Mass Spectrom., 2009, 23, 1825-1833.

H.J.Andreazza, T.Wang, C.J.Bagley, P.Hoffmann and J.H.Bowie.  Negative ion fragmentations of deprotonated peptides.  The unusual case of isoAsp: a joint experimental and theoretical study.  Rapid Commun. Mass Spectrom., 2009, 23, 1993-2002.

T.Wang and J.H.Bowie.  A theoretical study of the rearrangement processes of energised CCCB and CCCAl.  Phys. Chem. Chem. Phys., 2009, 11, 7553-7561.

P.J.Sherman, R.J.Jackway, E.Nicholson, I.F.Musgrave, P.Boontheung and J.H.Bowie.  Activities of seasonably variable caerulein and rothein peptides from the tree frogs Litoria splendida and Litoria rothii.  Toxicon, 2009, 54, 828-835.

D.Bilusich, R.J.Jackway, I.F.Musgrave, M.J.Tyler and J.H.Bowie.  The host-defence skin peptides of Peron's Tree Frog Litoria peronii in winter and summer.  Sequence determination by electrospray mass spectrometry and activities of the peptides.  Rapid Commun. Mass Spectrom., 2009, 23, 2628-2636.

T.Wang, M.A.Buntine and J.H.Bowie.  A theoretical study of the isomers of isoelectronic C4, (C3B)- and (C3N)+. Rearrangements through cyclic isomers. J. Phys. Chem. A, 2009, 113, in press.

M.J.Maclean, S.Walker, T.Wang, P.C.H.Eichinger, P.J.Sherman and J.H.Bowie.  Diagnostic fragmentations of adducts formed between carbanions and carbon disulfide. A joint experimental and theoretical study.  Org. Biomol. Chem., 2009, in press.

T.Wang and J.H.Bowie.  Gas phase intramolecular anion rearrangement of some trimethylsilyl containing systems revisited.  A theoretical approach. Rapid Commun. Mass Spectrom., 2010, 24, in press.

P.J.Sherman, R.J.Jackway, J.D.Gelman, S.Praporski, G.A.McCubbin, A.Mechler, L.L.Martin, F.Separovic and J.H.Bowie.  The solution structure and membrane interactions of the antimicrobial peptide fallaxidin 4.1a: an NMR and QCM study. Biochemistry, 2010, in press.

J.H.Bowie.  Splendipherin: the aquatic male sex pheromone of the Magnificent Tree Frog Litoria splendida. in Pheromones, Theories, Types and Uses. F.Columbus ed, Nova Publishers, New York, USA, 2010, in press.

P.J.Sherman, J.H.Bowie and M.J.Tyler. Divergence in populations of Ewing's Tree Frog, Litoria ewingi, in south eastern Australia.  The application of skin peptide profiling.   Aust. J. Zool., 2010, in press.

T.Wang, S.Dua and J.H.Bowie.  The formation of NCCCCCN by charge stripping of (NCCCCCN)-. in the gas phase.  How does energized NCCCCCN lose C?  A joint experimental and theoretical study,  J. Phys. Chem., 2010, in press.

 

Expertise for Media Contact

Categories	Science and technology
Expertise	Mass spectrometry Interstellar molecules Bioactive peptides from Australian animals
Notes	Alt phone: (08) 8303 5996

Entry last updated: Sunday, 22 Nov 2009