Structural analysis of bikunin glycosaminoglycan.
The structure of an intact glycosaminoglycan (GAG) chain of the bikunin proteoglycan (PG) was analyzed using a combined top-down and bottom-up sequencing strategy. PGs are proteins with one or more linear, high-molecular weight, sulfated GAG polysaccharides O-linked to serine or threonine residues. GAGs are often responsible for the biological functions of PGs, and subtle variations in the GAG structure have pronounced physiological effects. Bikunin is a serine protease inhibitor found in human amniotic fluid, plasma, and urine. Bikunin is posttranslationally modified with a chondroitin sulfate (CS) chain, O-linked to a serine residue of the core protein. Recent studies have shown that the CS chain of bikunin plays an important role in the physiological and pathological functions of this PG. While no PG or GAG has yet been sequenced, bikunin, the least complex PG, offers a compelling target. Electrospray ionization Fourier transform-ion cyclotron resonance mass spectrometry (ESI FTICR-MS) permitted the identification of several major components in the GAG mixture having molecular masses in a range of 5505-7102 Da. This is the first report of a mass spectrum of an intact GAG component of a PG. FTICR-MS analysis of a size-uniform fraction of bikunin GAG mixture obtained by preparative polyacrylamide gel electrophoresis, allowed the determination of chain length and number of sulfo groups in the intact GAGs.
Chi
Lianli
L
Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, Troy, New York 12180, USA.
Wolff
Jeremy J
JJ
Laremore
Tatiana N
TN
Restaino
Odile F
OF
Xie
Jin
J
Schiraldi
Chiara
C
Toida
Toshihiko
T
Amster
I Jonathan
IJ
Linhardt
Robert J
RJ
D000510:Alpha-Globulins / Q000737:chemistry*; D002240:Carbohydrate Sequence; D006025:Glycosaminoglycans / Q000737:chemistry*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011509:Proteoglycans / Q000737:chemistry*
D000510:Alpha-Globulins; D006025:Glycosaminoglycans; D011509:Proteoglycans; C016001:alpha-1-microglobulin