Rapid purification and properties of human glycodelin (endometrial alpha2-globulin).
The method presented can easily produce milligram amounts of glycodelin from pregnancy endometrium, with a 19% yield. It involves anion-exchange chromatography, gel permeation and chromatofocusing; it results in one stainable band at Mr 28,000 after sodium dodecyl sulphate-polyacrylamide electrophoresis, as well as after immunoblot analysis, performed using an affinity-purified IgG fraction from an antiserum against glycodelin. In spite of this, the corresponding gel isoelectric focusing pattern gives four stainable bands with pI values between 4.55 and 5.2. Western immunoblot analysis of tissue extracts indicates the presence of glycodelin epitopes associated with materials heavier than the native protein. Circular dichroism spectra of the highly purified protein in water solutions indicate a large amount of beta-sheet conformation, whereas those obtained with different proportions of 2-propanol in water, show an increased proportion of alpha-helix conformation.
Pala
A
A
Laboratory of Biochemistry of Sex Hormones of the First and Second Institute of Gynaecology and Obstetrics and Department of Chemistry, University La Sapienza, Rome, Italy.
Padula
F
F
Barteri
M
M
Benagiano
M
M
Gaudiano
M C
MC
Moro
M
M
Benagiano
G
G
D000838:Anions; D015153:Blotting, Western; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D004717:Endometrium / Q000737:chemistry*; D005260:Female; D000071816:Glycodelin; D006023:Glycoproteins / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D008970:Molecular Weight; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000737:chemistry / Q000302:isolation & purification*; D017433:Protein Structure, Secondary
D000838:Anions; D000071816:Glycodelin; D006023:Glycoproteins; C045246:PAEP protein, human; D011257:Pregnancy Proteins