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The proteoglycan bikunin has a defined sequence

Abstract

Proteoglycans are complex glycoconjugates that regulate critical biological pathways in all higher organisms. Bikunin, the simplest proteoglycan, with a single glycosaminoglycan chain, is a serine protease inhibitor used to treat acute pancreatitis. Unlike nucleic acids and proteins, whose synthesis is template driven, Golgi-synthesized glycosaminoglycans are not believed to have predictable or deterministic sequences. Bikunin peptidoglycosaminoglycans were prepared and fractionated to obtain a collection of size-similar and charge-similar chains. Fourier transform mass spectral analysis identified a small number of parent molecular ions corresponding to monocompositional peptidoglycosaminoglycans. Fragmentation using collision-induced dissociation unexpectedly afforded a single sequence for each monocompositional parent ion, unequivocally demonstrating the presence of a defined sequence. The biosynthetic pathway common to all proteoglycans suggests that even more structurally complex proteoglycans, such as heparan sulfate, may have defined sequences, requiring a readjustment in the understanding of information storage in complex glycans.

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Figure 1: Biosynthetic pathway for chondroitin sulfate A GAG.
Figure 2: FT-ICR-MS analysis of polydisperse and heterogeneous bikunin peptidoglycosaminoglycan.
Figure 3: FT-ICR-MS analysis of a bikunin peptidoglycosaminoglycan fraction.
Figure 4: Bikunin sequencing flow chart.

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Acknowledgements

The authors thank D. Zagorevski for his expertise in the proteomics core at Rensselaer Polytechnic Institute and the US National Institutes of Health for support (GM38060).

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M.L. and F.E.L. III contributed experiments and data interpretation. T.N.L. contributed the fractions for analysis and assisted in writing. T.T. contributed the bikunin and assisted in writing. R.J.L. and I.J.A. contributed experimental planning, result interpretation and wrote the paper.

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Correspondence to Robert J Linhardt.

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Ly, M., Leach, F., Laremore, T. et al. The proteoglycan bikunin has a defined sequence. Nat Chem Biol 7, 827–833 (2011). https://doi.org/10.1038/nchembio.673

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