Characterization of lectin aggregates in the hemolymph of freshwater prawn Macrobrachium rosenbergii.
In invertebrates, lectins play relevant roles in innate immunity; however, their regulatory mechanisms have not been identified yet. In this work, we purified, by gel filtration and affinity chromatography, lectin aggregates circulating in the hemolymph of the freshwater prawn Macrobrachium rosenbergii and compared their physicochemical properties with a previously described lectin (MrL). High-molecular weight MrL aggregates (MrL-I) lack hemagglutinating activity and showed bands of 62.1, 67.1 and 81.4 kDa, whereas MrL-III, which corresponds to MrL, showed hemagglutinating activity and is constituted by a single 9.6-kDa band as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. MrL-I and MrL-III showed similar amino acid composition but different carbohydrates concentration. Edman degradation indicated NH2-terminal sequence of five amino acids for the 9.6-kDa MrL-III (DVPLL/A) and eleven for the main 81.4-kDa band identified in MrL-I (DVPLL/AXKQQQD); analysis by MALDI-TOF indicated a different tryptic pattern for MrL-I and MrL-III. MrL-I was recognized by monoclonal antibodies against MrL-III. Circular dichroism indicated that the secondary structure in both proteins is similar and contains 23% of beta-sheet and 24% of alpha-helix. Our results suggest that differential posttranslational processes that favor aggregation are involved in regulating the activity of the lectin.
Departamento de Bioquímica, Instituto Nacional de Enfermedades Respiratorias, 10245, Mexico.
Ali
A
Pereyra
Roberto
R
Zenteno
Lorena
L
Vázquez
Salvador
S
Martínez-Cairo
Adela
A
Rodríguez
Guillermo
G
Mendoza-Hernández
Edgar
E
Zenteno
Concepción
C
Agundis
D000595:Amino Acid Sequence; D000818:Animals; D002846:Chromatography, Affinity; D002942:Circular Dichroism; D003445:Crustacea / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006458:Hemolymph / Q000378:metabolism*; D037102:Lectins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D020033:Protein Isoforms / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
D037102:Lectins; D020033:Protein Isoforms