Human colon sialidase: characterization and activity levels in normal mucosa and colonic adenocarcinoma.
Human colon sialidase has been characterized, and its activity levels in normal mucosa and colonic adenocarcinoma have been determined. Sialidase activity was maximal at pH 5.5, and was unstable with storage at 4 and -20 degrees C. The bulk of activity was pellet-associated, and could not be released with triton X-100 or 3-([3-cholamidopropyl]- dimethylammonio)-1-propanesulfonate. Using 2'-(4-methylumbelliferyl)alpha-D-N-acetylneuraminic acid as substrate, the Km and Vmax values were estimated to be 0.140 mmol/l and 63 mU/g, respectively. Furthermore, an inhibition by substrate concentrations above 1.5 mmol/l was detected. Neuraminic acid caused a competitive inhibition with a Ki of 3.5 mmol/l. A statistically significant increase (p < 0.001) in the sialidase specific activity was found in primary colonic adenocarcinoma (104.20 +/- 8.00 mU/g) compared to that of the normal mucosa (72.50 +/- 7.67 mU/g).
Martínez-Zorzano
V S
VS
Department of Fundamental Biology, University of Vigo, Spain.
Feijoo
C
C
Páez de la Cadena
M
M
Butrón
M
M
Fernández-Briera
A
A
Rodríguez-Berrocal
F J
FJ
D000230:Adenocarcinoma / Q000175:diagnosis / Q000201:enzymology*; D002413:Cations, Divalent / Q000737:chemistry; D002414:Cations, Monovalent / Q000737:chemistry; D002793:Cholic Acids / Q000737:chemistry; D003106:Colon / Q000201:enzymology*; D003110:Colonic Neoplasms / Q000175:diagnosis / Q000201:enzymology*; D003902:Detergents / Q000737:chemistry; D004795:Enzyme Stability; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006923:Hymecromone / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D007413:Intestinal Mucosa / Q000201:enzymology; D009439:Neuraminidase / Q000378:metabolism*; D017830:Octoxynol / Q000737:chemistry; D011506:Proteins / Q000032:analysis; D012995:Solubility; D013696:Temperature
D002413:Cations, Divalent; D002414:Cations, Monovalent; D002793:Cholic Acids; D003902:Detergents; D005456:Fluorescent Dyes; D011506:Proteins; D006923:Hymecromone; C021224:2'-(4-methylumbelliferyl)-alpha-D-N-acetylneuraminic acid; D017830:Octoxynol; D009439:Neuraminidase; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate