Abstract
In a prior study we showed evidence that latex from Carica candamarcensis contains a protein fraction that stimulates mammalian cell proliferation. In this report we describe the isolation of two proteinases responsible for this effect. Both proteinases (P1, P2) display a relative mass of 23 kDa and following chromatographic purification stimulate proliferation of fibroblastic and epithelial cells. P2 added to L929 fibroblasts at 2.5 nM enhances proliferation by 60 %. We further demonstrate that its cellular effect is linked to an increase in activity of Erk2, a component of the MAP kinase pathway. To our knowledge, this is the first known plant proteinase to exert a proliferative effect in mammalian cells. This novel mitogenic property attributed to a purified cysteine proteinase may explain some of the therapeutic actions attributed to these enzymes.
Key words
Carica candamarcensis
- Cariaceae - latex - mitogens - proteinases
References
-
1
Dussourd D E, Eisner T.
Vein-cutting behavior: insect counterploy to the latex defense of plants.
Science.
1987;
237
898-901
-
2
Silva L G, Garcia O, Lopes M TP, Salas C E.
Changes in protein profile during coagulation of latex from Carica papaya
.
Braz J Med Biol Res.
1997;
30
615-9
-
3
Furie B, Furie B C.
The molecular base of blood coagulation.
Cell.
1988;
53
505-18
-
4
Moutim V, Silva L G, Lopes M TP, Fernandes G W, Salas C E.
Spontaneous processing of peptides during coagulation of latex from Carica papaya
.
Plant Sci.
1999;
142
115-21
-
5 Clarck R AF. The molecular and cellular biology of wound repair. Second edition Editor Clarck RAF Plenum Press New York; 1998
-
6
Ahmed S U, Bar-Peled M, Raikhel N V.
Cloning and subcellular location of an Arabidopsis receptor-like protein that shares common features with protein-sorting receptors of eukaryotic cells.
Plant Physiol.
1997;
114
325-36
-
7
Komatsu S, Abe K, Karibe H.
Epidermal growth factor binding proteins in rice.
Biol Pharm Bull.
1996;
19
905-8
-
8
Kohorn B D, Lane S, Smith T A.
An Arabidopsis serine/threonine kinase homologue with an epidermal growth factor repeat selected in yeast selected for its specificity for a thylakoid membrane protein.
Proc Natl Acad Sci.
1992;
89
10 989-92
-
9
Molloy C J, Pawlowski J E, Taylor D S, Turner C E, Weber H, Peluso M.
Thrombin receptor activation elicits rapid protein tyrosine phosphorylation and stimulation of the raf-1/MAP kinase pathway preceding delayed mitogenesis in cultured rat aortic smooth muscle cells: evidence for an obligate autocrine mechanism promoting cell proliferation induced by G-protein-coupled receptor agonist.
J Clin Invest.
1996;
97
1173-83
-
10
Bravo L M, Hermosilla L, Salas C E.
A biochemical comparison between latex from C. candamarcensis and C. papaya
.
Braz J Med Biol Res.
1994;
27
2831-42
-
11
Baeza G, Correa D, Salas C E.
Proteolytic enzymes in C. candamarcensis
.
J Sci Food and Agric.
1990;
51
1-9
-
12
Silva C A, Gomes M TR, Ferreira R S, Rodrigues K CL, do Val C G, Lopes M TP, Mello V J, Salas C E.
A mitogenic protein fraction in latex from Carica candamarcensis
.
Planta Medica.
2003;
69
1-7
-
13
Gravina de Moraes M, Termignoni C, Salas C E.
Biochemical characterization of a new cysteine endopeptidase from Carica candamarcensis L.
Plant Sci.
1994;
102
11-18
-
14
Bradford M M.
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
Anal Biochem.
1976;
72
248-54
-
15
Pereira M T, Lopes M TP, Meira W O, Salas C E.
Purification of a cysteine proteinase from Carica candamarcensis L. and cloning of a genomic putative fragment coding for this enzyme.
Protein Express Purif.
2001;
22
249-57
-
16
Mynott T L, Ladhams A, Scarmato P, Engwerda C R.
Bromelain from pineapple stems, proteolitically blocks activation of extracellular regulated kinases-2 in T cells.
J Immunol.
1999;
163
2568-75
-
17
Houck J C, Chang C M, Klein G.
Isolation of an effective debriding agent from the stems of pineapple plants.
Int J Tissue React.
1983;
5
125-34
-
18
Hewitt H, Whittle S, Lopez S, Bailey E, Weaver S.
Topical use of papaya in chronic skin ulcer therapy in Jamaica.
West Indian Med J.
2000;
49
32-3
-
19
Starley I F, Mohammed P, Schneider G, Bickler S W.
The treatment of paediatric burns using topical papaya.
Burns.
1999;
25
636-9
-
20
Ford C N, Reinhard E R, Yeh D, Syrek D, De Las Morenas A, Bergman S B, Williams S, Hamori C A.
Interim analysis of a prospective, randomized trial of vacuum-assisted closure versus the healthpoint system in the management of pressure ulcers.
Ann Plast Surg.
2002;
49
55-61
-
21
Walreavens V, Jaziri M, van Beeumen J, Schnek A G, Kleinschmidt T, Looze Y.
Isolation and preliminary characterization of the cysteine-proteinases from the latex of Carica candamarcensis Hook.
Biol Chem Hoppe-Seyler.
1993;
374
501-6
Carlos E Salas, PhD
Instituto de Ciências Biológicas
Universidade Federal de Minas Gerais
Av Antônio Carlos 6627
Belo Horizonte
MG 31270-901
Brasil
Telefon: +55-31-3499-2646
Fax: +55-31-3499-2646
eMail: cesbufmg@mono.icb.ufmg.br
