The effect of the carbohydrate moiety upon the size and conformation of human plasma galactoglycoprotein as judged by electron microscopy and circular dichroism. Structural studies of a glycoprotein after stepwise enzymic carbohydrate removal.
Galactoglycoprotein is a unique human plasma protein [76% carbohydrate (23% N-acetylneuraminic acid, 20% galactose, 3% mannose, 1% fucose and 29% N-acetylgalactosamine plus N-acetylglucosamine) and 24% polypeptide, a single polypeptide chain of about 200 amino acid residues that is high in serine and threonine content] [Schmid, Mao, Kimura, Hayashi & Binette (1980) J. Biol. Chem. 255, 3221-3226]. Highly purified exoglycosidases with well-defined specificities were used to prepare five derivatives of galactoglycoprotein in which sequential residues of N-acetylneuraminic acid, galactose, N-acetylglucosamine, a second galactose and N-acetylgalactosamine were removed with 83% of the total carbohydrate cleaved. C.d. shows that native galactoglycoprotein and all derivatives in aqueous buffer are predominantly random coil, suggesting that removal of a large number of electrostatic net charges, as well as the major portion of the carbohydrate moiety, does not alter the secondary structure of the polypeptide chain. Examination of the size and conformation of tungsten-shadowed galactoglycoprotein and asialo and agalacto derivatives by electron microscopy shows the size and conformation of all three preparations to be similar, with only minor differences in particle length and width.
Watzlawick
H
H
Institut für Biochemie II, University of Heidelberg, Federal Republic of Germany.
Walsh
M T
MT
Ehrhard
I
I
Slayter
H S
HS
Haupt
H
H
Schwick
H G
HG
Jourdian
G W
GW
Hase
S
S
Schmid
K
K
Brossmer
R
R
D015703:Antigens, CD; D001798:Blood Proteins / Q000737:chemistry*; D002240:Carbohydrate Sequence; D002942:Circular Dichroism; D006023:Glycoproteins / Q000097:blood* / Q000737:chemistry*; D006026:Glycoside Hydrolases / Q000494:pharmacology; D006801:Humans; D051917:Leukosialin; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D011487:Protein Conformation; D012795:Sialoglycoproteins
D015703:Antigens, CD; D001798:Blood Proteins; D006023:Glycoproteins; D051917:Leukosialin; C465014:SPN protein, human; D012795:Sialoglycoproteins; D006026:Glycoside Hydrolases