Академический Документы
Профессиональный Документы
Культура Документы
Lehrstuhlfiir Pharmazeutische Biologie, Universitiit Miinchen, Karlstr. 29, D-8000 Miinchen 2 and *Genzentrum der
Universitiit Miinchen. Am Kloperspitz, D-8033 Martinsried, FRG
Exposure of several species of the order Fabales to Cd*+ results in the formation of metal chelating peptides
of the general structure (y-Glu-Cys),-/?-Ala (n = 2-7). They are assumed to be formed from homo-glutathi-
one and are termed homo-phytochelatins, as they are homologous to the recently discovered phytochelatins.
These peptides are induced by a number of metals such as CdZ+, Zn*+, HgZ+, Pb2+, AsOd2- and others.
They are assumed to detoxify poisonous heavy metals and to be involved in metal homeostasis.
48
Volume 205, number 1 FEBSLETTERS September 1986
teine. By cyclic repetition of the y-glutamyl of the peptide fraction. The M, of the largest h-PC
transferase and Edman degradation treatments, thus far observed is 1715.
the complete sequence of the peptide (peak 2) h-PCs are the heavy metal chelators in all the
could be elucidated. homo-glutathione containing plant species up to
The two minor peptides were individually now analysed. The members with increasing
dinitrophenylated and oxidized with performic molecular mass are present in decreasing concen-
acid. Partial acid hydrolysates of the modified tration as previously observed for the PC series
peptides were separated by two dimensional TLC [3,41.
revealing, respectively, a set of 6 and 8 partially The synthesis of h-PCs is induced in these plants
hydrolyzed products. Quantitative analysis of by heavy metal ions such as AsO:-, Cd?+, Ga3+,
these peptides afforded the expected dinitrophen- Zn’+, Hg’+, Pb2+, Ag+, Sb3+. No induction of h-
ylated peptide fragments. PC was recorded with A13+, B”+, Ba’+, Ca2+,
From these experiments it can be concluded that Ce3+, Cs+, K+, Mg2+, Mn’+.
the chain length pattern of these compounds is h-GSH has previously been discovered and
identical to the one observed in the phytochelatin characterized in P. aureus [8,9]. It is also present
series. The only difference, however, resides in the in mung bean [lo] and has been suggested to occur
carboxy-terminus where P-alanine substitutes for in G. max, P. limensis, P. vulgaris and Trifolium
glycine. The amino acid sequence and composition repens [ll]. The absence of glutathione (GSH)
of the soja-derived metal-binding peptides is given from the above mentioned Phaseoleae plants was
in the general formula depicted in fig.2. verified by HPLC analysis. Instead of glutathione
The term homo-phytochelatin (h-PC) is applied its @-alanine containing homologue, homo-
to this set of compounds containing ,&alanine, as glutathione (h-GSH; fig.2; n = 1) was isolated from
compared to the normal glycine containing each member of these plants and unequivocally
phytochelatins (PCs), which represent the prin- identified.
cipal heavy-metal-binding peptides in the plant Exposure of plants of the tribes Ononideae,
kingdom with the exception of the order Fabales Trifolieae, Vicieae, Astragaleae, and Loteae, all of
ill. the order Fabales, to Cd2+ revealed a peptide pat-
tern which is exemplified by Lathyrus ochrus
3.2. Distribution of homo-phytochelatins depicted in fig.lC. Clearly, double peaks could be
in higher plants perceived signifying the presence of both GSH and
Seedlings or cell suspension cultures of the order h-GSH (peak 1) as well as for PC2 (peak 2).
Fabales were exposed to 20 PM Cd(NO& for 4 Separate analysis confirmed the assumption that in
days, and the exclusive formation of h-PCs was these plants both GSH and its&alanine containing
observed in the following plants: Phaseolus homologue (h-GSH) co-occur. This pattern is also
vulgaris, P. coccineus, P. aureus, P. lunatus, P. applicable to the metal induced chelating peptides.
multifloris, Canavalia ensiformis, Cajanus cajan, Both sets of PC as well as h-PC are induced by
Dolichos lablab, G. max, G. clandestina, Cd2+ in these plants. The following species of the
Erythrina crista-galli, E. melanacantha, E. cor- order Fabales so far analysed revealed this mixed
alloides, Rhynchosia phaseoloides. As an example pattern: Astragalus lusitanicus, A. cicer, A. gum-
fig.lB illustrates that the exposure of P. aureus to mifer, Cicer arietinum, Coronilla varia, Galega of-
Cd(NO& induces h-PCs with up to seven (y-Glu- ficinalis, L. ochrus, L. odoratus, L. silvester, Lens
Cys) units (h-PCT), as seen from the HPLC pattern culinaris, Lotus ornithopodioides, Melilotus alba,
Ononis natrix, Trifolium pratense, T. incarnatum,
T. badium, T. subterraneum, T. lupinaster, T.
pannonicum, Trigonella coerulea, T.
foenum-graecum.
49
Volume 205, number 1 FEBS LETTERS September 1986
of heavy metals exclusively through the formation fact that is displayed by the ability of these plants
of peptides which consist of a linear chain of y- to synthesize both PCs as well as h-PCs upon ex-
bound L-glutamic acid, L-cysteine and a carboxy- posure to heavy metals. The function of the PC
terminal fl-alanine only. These compounds were and h-PC peptide series is apparently assigned to
termed homo-phytochelatins. detoxification and homeostasis of heavy metals in
Because of the y-glutamyl linkages and presence plants.
of ,&alanine, for which no triplet code exists, the
h-PCs are not synthesized via mRNA and
ACKNOWLEDGEMENTS
therefore cannot be regarded as primary gene pro-
ducts. The exclusive formation of h-PCs in h-GSH This study was supported by Bundesminister fur
containing Fabales and the co-occurrence of pep- Forschung und Technologie, Bonn, and Fonds der
tides containing either glycine or P-alanine at the Chemischen Industrie. The excellent technical
carboxy-terminus within the same plant species are assistance of Miss Susanne Kunz is gratefully
indicative that the biosynthesis of PCs or h-PCs acknowledged.
occurs by the consumption of GSH or h-GSH. As
previously suggested for GSH, the tripeptide and
REFERENCES
its homologue seem to be the substrates for PC
synthesis [1,3,4]. The occurrence of GSH or h- 111Grill, E., Winnacker, E.-L. and Zenk, M.H. (1985)
GSH in plants would thus determine PC or h-PC Science 230, 674-675.
formation. [21 Kondo, N., Isobe, M., Imai, K. and Goto, T.
Analysis of plants from the primitive algae to (1985) Agric. Biol. Chem. 49, 71-83.
the orchids as to their abilities to chelate heavy 131Grill, E., Winnacker, E.-L. and Zenk, M.H. (1986)
FEBS Lett. 197, 115-120.
metals revealed that algae, ferns, gymnospermae
and angiospermae have developed the mechanism 141Grill, E., Winnacker, E.-L. and Zenk, M.H. (1986)
submitted.
to sequester heavy metals by PC formation. Other
151Hoagland, D.R. and Snijder, W.C. (1933) Proc.
mechanisms have not been identified [ 11. The ubi- Am. Sot. Hortic. Sci. 30, 288-296.
quity of PC as the heavy metal binding principle in 161Orlowski, M. and Meister, A. (1965) J. Biol.
plants is only limited by the presence of its Chem. 240, 338-347.
homologue h-PC in some higher plants of the [71 Chang, J.Y. (1981) Biochem. J. 199, 557-564.
order Fabales, reflecting the complete or partial 181Carnegie, P.R. (1963) Biochem. J. 89, 459-471.
substitution of GSH by h-GSH in these plants. The 191Carnegie, P.R. (1963) Biochem. J. 89, 471-478.
tribe Phaseoleae exclusively contains homo- UOI Magnicol, P.K. and Bergmann, L. (1984) Plant Sci.
glutathione. Therefore, h-PCs most probably Lett. 36, 219-223.
represent the Cd-binding components noticed Ufl Price, C.A. (1957) Nature 180, 148-149.
1121Casterline, J.L. and Barnett, N.M. (1982) Plant
earlier to occur in soybean [12] and P. vulgaris Physiol. 69, 1004-1007.
[ 131. Other tribes of the Fabales seem to be tran- 1131Weigel, H.J. and Jlger, H.J. (1980) Plant Physiol.
sient in that they contain both GSH and h-GSH, a 65, 480-482.
50