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Cited by 901Proteoglycans and glycosaminoglycans researchPolysaccharides and Plant Cell WallsCarbohydrate Chemistry and Synthesis

Three isoforms of mammalian hyaluronan synthases have distinct enzymatic properties

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Itano, Naoki, Sawai, Takahiro, Yoshida, Mamoru, Lenas, Petros, Yamada, Yoichi, Imagawa, Michiko, Shinomura, Tamayuki, Hamaguchi, Michinari +6 more·1999·Journal of Biological Chemistry
DOI 10.1074/jbc.274.35.25085
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Abstract

Three mammalian hyaluronan synthase genes, HAS1, HAS2, and HAS3, have recently been cloned. In this study, we characterized and compared the enzymatic properties of these three HAS proteins. Expression of any of these genes in COS-1 cells or rat 3Y1 fibroblasts yielded de novo formation of a hyaluronan coat. The pericellular coats formed by HAS1 transfectants were significantly smaller than those formed by HAS2 or HAS3 transfectants. Kinetic studies of these enzymes in the membrane fractions isolated from HAS transfectants demonstrated that HAS proteins are distinct from each other in enzyme stability, elongation rate of HA, and apparent K(m) values for the two substrates UDP-GlcNAc and UDP-GlcUA. Analysis of the size distributions of hyaluronan generated in vitro by the recombinant proteins demonstrated that HAS3 synthesized hyaluronan with a molecular mass of 1 x 10(5) to 1 x 10(6) Da, shorter than those synthesized by HAS1 and HAS2 which have molecular masses of 2 x 10(5) to approximately 2 x 10(6) Da. Furthermore, comparisons of hyaluronan secreted into the culture media by stable HAS transfectants showed that HAS1 and HAS3 generated hyaluronan with broad size distributions (molecular masses of 2 x 10(5) to approximately 2 x 10(6) Da), whereas HAS2 generated hyaluronan with a broad but extremely large size (average molecular mass of >2 x 10(6) Da). The occurrence of three HAS isoforms with such distinct enzymatic characteristics may provide the cells with flexibility in the control of hyaluronan biosynthesis and functions.

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  • Hyaluronan is synthesized by hyaluronan synthases located in the plasma membrane, and three isoforms synthesize hyaluronan chains of different lengths and at different speeds.

    “Hyaluronan is synthesized by hyaluronan sythases (HAS), which are located in the plasma membrane.Three isoforms of HAS exist (HAS-1, -2 and -3), encoded by individual genes, which synthesize hyaluronan chains of various length and at different speed, with HAS-1 and -2 generating hyaluronan chains of up to 2000 kD at fast speed, and HAS-3 producing hyaluronan shorter chains at slow speed (Itano et”

    From The hyaluronan and proteoglycan link proteins: Organizers of the brain extracellular matrix and key molecules for neuronal function and plasticity (2015) · cites this paper as Itano et al., 1999

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Authors
Itano, N., Sawai, T., Yoshida, M., Lenas, P., Yamada, Y., Imagawa, M., Shinomura, T., Hamaguchi, M., Yoshida, Y., Ohnuki, Y., Miyauchi, S., Spicer, A. P., McDonald, J. A., & Kimata, K.
Affiliations
  • Institute for Molecular Science of Medicine, Aichi Medical University (JP)
  • Nagoya University (JP)
  • Seikagaku Corporation (Japan) (JP)
  • University of California, Davis (US)
  • Mayo Clinic (US)
Corresponding author
Kimata, Koji
ORCID
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
1999-08
Volume
274
Issue
35
Pages
25085-25092
ISSN
0021-9258 (ISSN-L)
DOI
10.1074/jbc.274.35.25085
OpenAlex
W2167704983
Cited by
901
References
39
Citations by year
2026: 15 · 2025: 27 · 2024: 24 · 2023: 39 · 2022: 30 · 2021: 51 · 2020: 37 · 2019: 34 · 2018: 40 · 2017: 28 · 2016: 39 · 2015: 44 · 2014: 51 · 2013: 42 · 2012: 20
FWCI
7.45
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
National Institute of Arthritis and Musculoskeletal and Skin Diseases (1RO1AR44698)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Proteoglycans and glycosaminoglycans research
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Polysaccharides and Plant Cell Walls
  • Physical Sciences › Chemistry › Organic Chemistry › Carbohydrate Chemistry and Synthesis
Keywords
hyaluronan synthase, HAS1, HAS2, HAS3, hyaluronan synthesis, enzyme kinetic properties, UDP-GlcNAc, recombinant enzyme expression
MeSH terms
Hyaluronan Synthases, Animals, Cell Line, Enzyme Stability, Hyaluronic Acid, Isoenzymes, Kinetics, Membrane Proteins, Microscopy, Phase-Contrast, Recombinant Proteins, Substrate Specificity, Transfection, Transferases, Glucuronosyltransferase, Uridine Diphosphate Glucuronic Acid, Uridine Diphosphate N-Acetylglucosamine, Gene Expression, Glycosyltransferases, Xenopus Proteins

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