Purification and characterization of parvalbumins from silver carp (Hypophthalmichthy molitrix).
BACKGROUND
As the largest producer and consumer of freshwater fish in the world, many people suffer from allergy for consuming freshwater fish in China. However, the allergen profiles of freshwater fish are rarely known.


RESULTS
Parvalbumins (PVs) from the white muscle of silver carp (Hypophthalmichthy molitrix) were purified by ammonium sulfate fractionation and column chromatography including DEAE-Sepharose and Superdex 75. Three PV isoforms-PV-I, PV-II, and PV-III-were obtained and their molecular masses as estimated by tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis were 12, 11, and 14 kDa, respectively. All the PVs could be detected by anti-frog PV monoclonal antibody. PV-I and PV-II were quite possibly glycoproteins, while PV-III was not glycosylated, as analyzed by periodic acid-Schiff (PAS) staining. Thermal stability revealed that PV-I and PV-II easily formed polymers, while these proteins were stable in a pH range of 4.0-10.0. A PV gene encoding 110 amino acid residues was cloned and it revealed high identity with PVs from other species of fish.


CONCLUSIONS
Three isotypes of PV were purified to homogeneity and one distinct PV gene was cloned in silver carp white muscle.
Liu
Guang-Ming
GM
College of Biological Engineering, Key Laboratory of Science and Technology for Aquaculture and Food Safety, Jimei University, Xiamen 361021, China.
Wang
Ning
N
Cai
Qiu-Feng
QF
Li
Teng
T
Sun
Le-Chang
LC
Su
Wen-Jin
WJ
Cao
Min-Jie
MJ
D000485:Allergens / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D002347:Carps / Q000235:genetics; D003001:Cloning, Molecular; D029941:Fish Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D006031:Glycosylation; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000737:chemistry*; D010320:Parvalbumins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D011108:Polymers; D020033:Protein Isoforms; D011898:Ranidae
D000485:Allergens; D000911:Antibodies, Monoclonal; D029941:Fish Proteins; D010320:Parvalbumins; D011108:Polymers; D020033:Protein Isoforms