Purification and biochemical characterization of Eumiliin from Euphorbia milii var. hislopii latex

被引：39

作者：

Fonseca, K. C. ^{[1
]}

Morais, N. C. G. ^{[1
]}

Queiroz, M. R. ^{[1
]}

Silva, M. C. ^{[1
]}

Gomes, M. S. ^{[1
]}

Costa, J. O. ^{[2
,3
]}

Mamede, C. C. N. ^{[1
]}

Torres, F. S. ^{[4
]}

Penha-Silva, N. ^{[2
]}

Beletti, M. E. ^{[1
]}

Canabrava, H. A. N. ^{[1
]}

Oliveira, F. ^{[1
]}

机构：

[1] Univ Fed Uberlandia, Inst Ciencias Biomed, BR-38400 Uberlandia, MG, Brazil

[2] Univ Fed Uberlandia, Inst Genet & Bioquim, BR-38400 Uberlandia, MG, Brazil

[3] Inst Fed Educ Ciencias & Tecnol Triangulo Mineiro, Campus Ituiutaba, MG, Brazil

[4] Univ Fed Minas Gerais, Inst Ciencias Biol, Belo Horizonte, MG, Brazil

来源：

PHYTOCHEMISTRY | 2010年 / 71卷 / 07期

关键词：

Euphorbia milii; Crown-of-Thorns; Euphorbiaceae; Cysteine protease; Eumiliin; CUCUMISIN-LIKE PROTEASE; THROMBIN-LIKE ENZYME; SERINE-PROTEASE; ENDEMIC AREA; ANTIPLATELET; PROTEINASE; COAGULANT; SARCOCARP; CLONING; VENOMS;

D O I：

10.1016/j.phytochem.2010.02.009

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A protease, which we designate Eumiliin, was isolated from the latex of Euphorbia milii var. hislopii by a combination of ion-exchange chromatographic steps using DEAE-Sephacel and gel-filtration with Sephadex G-75. Eumiliin is a monomeric protein with an apparent molecular mass of 30 kDa by SDS-PAGE under reducing conditions and gave one main peak at 29,814 KDa in MALDI-TOF/TOF mass spectrometry. Eumiliin has caseinolytic and fibrinogenolytic activities, but no hemorrhagic or defibrinating activities. The enzyme readily hydrolyzes the A alpha-chain of fibrinogen and, more slowly, the B beta-chain. Its fibrinogenolytic activity is inhibited by beta-mercaptoethanol and leupeptin. In contrast, EDTA and benzamidine did not affect the activity of Eumiliin. The caseinolytic activity of Eumiliin had a pH optimum of 8.0 and was stable in solution at up to 40 degrees C; activity was completely lost at >= 80 degrees C. Intraplantar injection of Eumiliin (1-25 mu g/paw) caused a dose- and time-dependent hyperalgesia, which peaked 1-5 h after enzyme injection. Intraplantar injection of Eumiliin (1-25 mu g/paw) also caused an oedematogenic response that was maximal after 1 h. Morphological analyses indicated that Eumiliin induced an intense myonecrosis, with visible leukocyte infiltrate and damaged muscle cells 24 h after injection. (C) 2010 Elsevier Ltd. All rights reserved.

引用

页码：708 / 715

页数：8

共 52 条

[1]

[Anonymous], 1957, ARCH I PHARMACODYN

[2] Isolation and characterization of a serine protease from the sprouts of Pleioblastus hindsii Nakai [J].

Arima, K ;

Uchikoba, T ;

Yonezawa, H ;

Shimada, M ;

Kaneda, M .

PHYTOCHEMISTRY, 2000, 54 (06) :559-565

[3] Cucumisin-like protease from the latex of Euphorbia supina [J].

Arima, K ;

Uchikoba, T ;

Yonezawa, H ;

Shimada, M ;

Kaneda, M .

PHYTOCHEMISTRY, 2000, 53 (06) :639-644

[4] EVALUATION OF THE MOLLUSCICIDAL PROPERTY OF EUPHORBIA-SPLENDENS VAR HISLOPII (NEB) (EUPHORBIACEAE) .2. INVESTIGATION IN LOTIC HABITAT [J].

BAPTISTA, DF ;

VASCONCELLOS, MC ;

LOPES, FEF ;

SILVA, IP ;

SCHALL, VT .

MEMORIAS DO INSTITUTO OSWALDO CRUZ, 1992, 87 (04) :549-553

[5]

BARRIE WW, 1976, ARCH SURG-CHICAGO, V5, P561

[6] Cytotoxic and antiviral activities of Colombian medicinal plant extracts of the Euphorbia genus [J].

Betancur-Galvis, LA ;

Morales, GE ;

Forero, JE ;

Roldan, J .

MEMORIAS DO INSTITUTO OSWALDO CRUZ, 2002, 97 (04) :541-546

[7]

Bhatnagar V. P., 2000, Journal of Medicinal and Aromatic Plant Sciences, V22-23, P464

[8]

CHANG MC, 1995, J LAB CLIN MED, V125, P508

[9] An unusual thermostable aspartic protease from the latex of Ficus racemosa (L.) [J].

Devaraj, K. B. ;

Gowda, Lalitha R. ;

Prakash, V. .

PHYTOCHEMISTRY, 2008, 69 (03) :647-655

[10]

Domingos A, 2000, EUR J BIOCHEM, V267, P6824, DOI 10.1046/j.1432-1327.2000.01780.x

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