Conformation of nucleoplasmin and its interaction with DNA-protamine complex as a simple model of fish sperm nuclei.
Nucleoplasmin was isolated from Xenopus laevis eggs and purified by an improved method using an open column. Its conformation was investigated spectrophotometrically by UV, CD and fluorescence. It was shown that alpha-helix content of nucleoplasmin was 30-40%, and one of the two tryptophan residues in nucleoplasmin located in the hydrophobic surroundings and the other in the relatively hydrophilic surroundings. The isolated nucleoplasmin was found to decondense sperm nuclei of salmon also, suggesting a possibility of the existence of nucleoplasmin-like protein in fish as well. Collapse of the protamine (salmine)-DNA complex as a simple model for fish sperm nuclei by nucleoplasmin was directly observed by measuring OD320 of aqueous protamine-DNA mixtures. This is a molecular level observation for the removal of protamine from DNA-protamine complex.
Iwata
K
K
Division of Bioscience, Graduate School of Environmental Earth Science, Hokkaido University, Sapporo, Japan.
Hozumi
K
K
Itoh
T
T
Sakairi
N
N
Tokura
S
S
Katagiri
C
C
Nishi
N
N
D000595:Amino Acid Sequence; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism* / Q000648:ultrastructure; D002942:Circular Dichroism; D004247:DNA / Q000378:metabolism*; D005260:Female; D005453:Fluorescence; D008297:Male; D008969:Molecular Sequence Data; D009391:Nephelometry and Turbidimetry; D009687:Nuclear Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D056489:Nucleoplasmins; D010063:Ovum / Q000737:chemistry; D010750:Phosphoproteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D011479:Protamines / Q000378:metabolism*; D011487:Protein Conformation; D012473:Salmine / Q000378:metabolism; D012474:Salmon; D013053:Spectrophotometry / Q000379:methods; D013056:Spectrophotometry, Ultraviolet; D013094:Spermatozoa / Q000737:chemistry / Q000502:physiology*; D014982:Xenopus laevis
D009687:Nuclear Proteins; D056489:Nucleoplasmins; D010750:Phosphoproteins; D011479:Protamines; D004247:DNA; D012473:Salmine