Purification, crystallization and preliminary X-ray analysis of CMS1MS2: a cysteine proteinase from Carica candamarcensis latex.
Cysteine proteinases from the latex of plants of the family Caricaceae are widely used industrially as well as in pharmaceutical preparations. In the present work, a 23 kDa cysteine proteinase from Carica candamarcensis latex (designated CMS1MS2) was purified for crystallization using three chromatography steps. The enzyme shows about fourfold higher activity than papain with BAPNA as substrate. Crystals suitable for X-ray diffraction experiments were obtained by the hanging-drop method in the presence of PEG and ammonium sulfate as precipitants. The crystals are monoclinic (space group P2(1)), with unit-cell parameters a = 53.26, b = 75.71, c = 53.23 A, beta = 96.81 degrees , and diffract X-rays to 1.8 A resolution.
Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antônio Carlos 6627, Belo Horizonte, MG, CEP 31270-901, Brazil.
Marco Túlio Ribeiro
MT
Gomes
Raphael Dias
RD
Teixeira
Henrique de Assis Lopes
Hde A
Ribeiro
Andréia Pereira
AP
Turchetti
Caroline Furtado
CF
Junqueira
Míriam Tereza Paz
MT
Lopes
Carlos Edmundo
CE
Salas
Ronaldo Alves Pinto
RA
Nagem
D001586:Benzoylarginine Nitroanilide / Q000494:pharmacology; D029441:Carica / Q000201:enzymology*; D002863:Chromogenic Compounds / Q000494:pharmacology; D018360:Crystallography, X-Ray; D003546:Cysteine Endopeptidases / Q000032:analysis* / Q000302:isolation & purification*; D007840:Latex / Q000737:chemistry*; D008970:Molecular Weight; D010940:Plant Proteins / Q000032:analysis* / Q000302:isolation & purification*; D014961:X-Ray Diffraction
D002863:Chromogenic Compounds; D007840:Latex; D010940:Plant Proteins; D001586:Benzoylarginine Nitroanilide; D003546:Cysteine Endopeptidases