Combining functional metagenomics and glycoanalytics to identify enzymes that facilitate structural characterization of sulfated N-glycans

被引:11
作者
Chuzel, Lea [1 ,2 ]
Fossa, Samantha L. [2 ]
Boisvert, Madison L. [2 ]
Cajic, Samanta [1 ]
Hennig, Rene [3 ]
Ganatra, Mehul B. [2 ]
Reichl, Udo [1 ,4 ]
Rapp, Erdmann [1 ,3 ]
Taron, Christopher H. [2 ]
机构
[1] Max Planck Inst Dynam Complex Tech Syst, Bioproc Engn, D-39106 Magdeburg, Germany
[2] New England Biolabs Inc, Ipswich, MA 01938 USA
[3] GlyXera GmbH, D-39120 Magdeburg, Germany
[4] Otto Von Guericke Univ, Chair Bioproc Engn, D-39106 Magdeburg, Germany
关键词
Sulfatase; Functional metagenomics; Glycoanalytics; Glycan analysis; Human microbiome; Glycan sulfation; N-acetylglucosamine-6-sulfate; PROTEIN GLYCOSYLATION; LINKED GLYCANS; IDENTIFICATION; OLIGOSACCHARIDES; CELLS; ROLES; SULFOGLYCOSIDASE; RIBOFLAVIN; THROUGHPUT; BIOMARKERS;
D O I
10.1186/s12934-021-01652-w
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Background Sulfate modification of N-glycans is important for several biological functions such as clearance of pituitary hormones or immunoregulation. Yet, the prevalence of this N-glycan modification and its functions remain largely unexplored. Characterization of N-glycans bearing sulfate modifications is hampered in part by a lack of enzymes that enable site-specific detection of N-glycan sulfation. In this study, we used functional metagenomic screening to identify enzymes that act upon sulfated N-acetylglucosamine (GlcNAc). Using multiplexed capillary gel electrophoresis with laser-induced fluorescence detection (xCGE-LIF) -based glycoanalysis we proved their ability to act upon GlcNAc-6-SO4 on N-glycans. Results Our screen identified a sugar-specific sulfatase that specifically removes sulfate from GlcNAc-6-SO4 when it is in a terminal position on an N-glycan. Additionally, in the absence of calcium, this sulfatase binds to the sulfated glycan but does not remove the sulfate group, suggesting it could be used for selective isolation of sulfated N-glycans. Further, we describe isolation of a sulfate-dependent hexosaminidase that removes intact GlcNAc-6-SO4 (but not asulfated GlcNAc) from a terminal position on N-glycans. Finally, the use of these enzymes to detect the presence of sulfated N-glycans by xCGE-LIF is demonstrated. Conclusion The present study demonstrates the feasibility of using functional metagenomic screening combined with glycoanalytics to discover enzymes that act upon chemical modifications of glycans. The discovered enzymes represent new specificities that can help resolve the presence of GlcNAc-6-SO4 in N-glycan structural analyses.
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页数:17
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共 69 条
  • [1] LECTIN-BASED GLYCOPROTEOMIC TECHNIQUES FOR THE ENRICHMENT AND IDENTIFICATION OF POTENTIAL BIOMARKERS
    Abbott, Karen L.
    Pierce, J. Michael
    [J]. METHODS IN ENZYMOLOGY, VOL 480: GLYCOBIOLOGY, 2010, 480 : 461 - 476
  • [2] Overlapping riboflavin supply pathways in bacteria
    Antonio Garcia-Angulo, Victor
    [J]. CRITICAL REVIEWS IN MICROBIOLOGY, 2017, 43 (02) : 196 - 209
  • [3] On the frequency of protein glycosylation, as deduced from analysis of the SWISS-PROT database
    Apweiler, R
    Hermjakob, H
    Sharon, N
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, 1999, 1473 (01): : 4 - 8
  • [4] The impact of glycosylation on the biological function and structure of human immunoglobulins
    Arnold, James N.
    Wormald, Mark R.
    Sim, Robert B.
    Rudd, Pauline M.
    Dwek, Raymond A.
    [J]. ANNUAL REVIEW OF IMMUNOLOGY, 2007, 25 : 21 - 50
  • [5] Matching the Diversity of Sulfated Biomolecules: Creation of a Classification Database for Sulfatases Reflecting Their Substrate Specificity
    Barbeyron, Tristan
    Brillet-Gueguen, Loraine
    Carre, Wilfrid
    Carriere, Cathelene
    Caron, Christophe
    Czjzek, Mirjam
    Hoebeke, Mark
    Michel, Gurvan
    [J]. PLOS ONE, 2016, 11 (10):
  • [6] Anaerobic sulfatase-maturating enzymes: Radical SAM enzymes able to catalyze in vitro sulfatase post-translational modification
    Benjdia, Alhosna
    Leprince, Jerome
    Guillot, Alain
    Vaudry, Hubert
    Rabot, Sylvie
    Berteau, Olivier
    [J]. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2007, 129 (12) : 3462 - +
  • [7] HUMAN UROKINASE CONTAINS GALNAC-BETA(1-4)[FUC-ALPHA(1-3)]GLCNAC-BETA(1-2) AS A NOVEL TERMINAL ELEMENT IN N-LINKED CARBOHYDRATE CHAINS
    BERGWERFF, AA
    THOMASOATES, JE
    VANOOSTRUM, J
    KAMERLING, JP
    VLIEGENTHART, JFG
    [J]. FEBS LETTERS, 1992, 314 (03) : 389 - 394
  • [8] Structure of a human lysosomal sulfatase
    Bond, CS
    Clements, PR
    Ashby, SJ
    Collyer, CA
    Harrop, SJ
    Hopwood, JJ
    Guss, JM
    [J]. STRUCTURE, 1997, 5 (02) : 277 - 289
  • [9] GlycoBase and autoGU: tools for HPLC-based glycan analysis
    Campbell, Matthew P.
    Royle, Louise
    Radcliffe, Catherine M.
    Dwek, Raymond A.
    Rudd, Pauline M.
    [J]. BIOINFORMATICS, 2008, 24 (09) : 1214 - 1216
  • [10] Function and structure of a prokaryotic formylglycine-generating enzyme
    Carlson, Brian L.
    Ballister, Edward R.
    Skordalakes, Emmanuel
    King, David S.
    Breidenbach, Mark A.
    Gilmore, Sarah A.
    Berger, James M.
    Bertozzi, Carolyn R.
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2008, 283 (29) : 20117 - 20125