scholarly journals Increased Expression of CD44 and Hyaluronate Synthase 3 Is Associated with Accumulation of Hyaluronate in Spongiotic Epidermis

2012 ◽  
Vol 132 (3) ◽  
pp. 736-738 ◽  
Author(s):  
Laurent Barnes ◽  
Pierre Carraux ◽  
Jean-Hilaire Saurat ◽  
Gürkan Kaya
Keyword(s):  
Hyaluronate Synthase

scholarly journals The hyaluronate synthase from a eukaryotic cell line

1993 ◽  
Vol 290 (3) ◽  
pp. 791-795 ◽  
Author(s):  
L Klewes ◽  
E A Turley ◽  
P Prehm
Keyword(s):  
Monoclonal Antibodies ◽  
Phase Separation ◽  
Affinity Chromatography ◽  
Cell Line ◽  
Aqueous Phase ◽  
Plasma Membranes ◽  
Eukaryotic Cell ◽  
Molecular Masses ◽  
Triton X ◽  
Hyaluronate Synthase

The hyaluronate synthase complex was identified in plasma membranes from B6 cells. It contained two subunits of molecular masses 52 kDa and 60 kDa which bound the precursor UDP-GlcA in digitonin solution and partitioned into the aqueous phase, together with nascent hyaluronate upon Triton X-114 phase separation. The 52 kDa protein cross-reacted with poly- and monoclonal antibodies raised against the streptococcal hyaluronate synthase and the 60 kDa protein was recognized by monoclonal antibodies raised against a hyaluronate receptor. The 52 kDa protein was purified to homogeneity by affinity chromatography with monoclonal anti-hyaluronate synthase.

scholarly journals Shedding of hyaluronate synthase from streptococci

1990 ◽  
Vol 267 (1) ◽  
pp. 191-196 ◽  
Author(s):  
A Mausolf ◽  
J Jungmann ◽  
H Robenek ◽  
P Prehm
Keyword(s):  
Membrane Protein ◽  
Protease Inhibitors ◽  
Chain Length ◽  
Sedimentation Rate ◽  
Culture Medium ◽  
Integral Membrane Protein ◽  
Phospholipid Vesicles ◽  
Free Solution ◽  
Hyaluronate Synthase

Hyaluronate synthase was shed into the culture medium from growing streptococci (group C) together with nascent hyaluronate. The mechanism of solubilization was analysed using isolated protoplast membranes. Solubilization increased when membranes were suspended in larger volumes, but it was temperature-independent and was not inhibited by protease inhibitors. Increased hyaluronate chain length enhanced solubilization. The soluble synthase could re-integrate into Streptococcal membranes in a saturable manner. The soluble synthase behaved like an integral membrane protein, although it was not integrated into phospholipid vesicles. In sucrose velocity centrifugation the synthase had a higher sedimentation rate in detergent-free solution, indicating that it existed in an aggregated state.

Expression of hyaluronate and hyaluronate synthase in human primary tumours and their metastases in scid mice

2002 ◽  
Vol 188 (1-2) ◽  
pp. 181-189 ◽  
Author(s):  
Milan Jojovic ◽  
Bertrand Delpech ◽  
Peter Prehm ◽  
Udo Schumacher
Keyword(s):  
Scid Mice ◽  
Hyaluronate Synthase

scholarly journals Hyaluronate synthase: cloning and sequencing of the gene from Streptococcus sp

1993 ◽  
Vol 289 (1) ◽  
pp. 179-184 ◽  
Author(s):  
M Lansing ◽  
S Lellig ◽  
A Mausolf ◽  
I Martini ◽  
F Crescenzi ◽  
...  
Keyword(s):  
Gene Locus ◽  
Southern Blotting ◽  
Peptide Binding ◽  
Predicted Amino Acid Sequence ◽  
Cloning And Sequencing ◽  
Reading Frame ◽  
Ribosome Binding ◽  
Translation Initiation Codon ◽  
Hyaluronate Synthase ◽  
Flanking Regions

The complete nucleotide sequence of hyaluronate synthase from Streptococcus sp. and its flanking regions is presented. The gene locus was designated has. Southern-blotting results suggested that the gene was conserved in hyaluronate-producing streptococci. A putative translation-initiation codon was identified and the open reading frame consists of 1566 bp, specifying a protein of 56 kDa. Sequences resembling the promoter and ribosome-binding site of Gram-positive organisms are found upstream of the synthase. The predicted amino-acid sequence reveals the presence of a 35-residue signal peptide. The sequence has some similarity to bacterial peptide-binding proteins.

scholarly journals Characterization of a high-Mr plasma-membrane-bound protein and assessment of its role as a constituent of hyaluronate synthase complex

1986 ◽  
Vol 237 (2) ◽  
pp. 343-357 ◽  
Author(s):  
N Mian
Keyword(s):  
Plasma Membranes ◽  
Kinase Activity ◽  
Glucuronic Acid ◽  
Quaternary Structure ◽  
Protein Molecule ◽  
Stationary Growth Phase ◽  
Membrane Bound ◽  
Hyaluronate Synthase

A high-Mr phosphoprotein (Mr 442,000) was purified from Nonidet-P-40-solubilized plasma membranes of cultured human skin fibroblasts. The protein comprised one 200,000-Mr subunit consisting of 116,000- and 84,000-Mr polypeptides and two identical 121,000-Mr subunits each consisting of 66,000- and 55,000-Mr polypeptides. The 200,000-Mr subunit and its polypeptides contained phosphotyrosine residues and were also [32P]phosphorylated at these residues from [gamma-32P]ATP in vitro by an intrinsic tyrosine kinase activity of the protein molecule in response to the presence of hyaluronate precursors, UDP-glucuronic acid and UDP-N-acetylglucosamine. The 121,000-Mr subunits and their polypeptides contained phosphoserine residues that could not be [32P]phosphorylated during autophosphorylation of the protein in vitro. The protein molecules separated from exponential- and stationary-growth-phase cells were identical in their quaternary structure, but appeared to exist in different proportions with respect to the state of phosphorylation of their 121,000-Mr subunits during different growth phases of the cell. Phosphorylation of polypeptides appeared to predispose in favour of their UDP-glucuronic acid- and UDP-N-acetylglucosamine-binding activities. The phosphorylated 116,000- and 84,000-Mr polypeptides of 200,000-Mr subunits possessed a single binding site for UDP-glucuronic acid and UDP-N-acetylglucosamine respectively. The phosphorylated 200,000-Mr subunit could also cleave the UDP moiety from UDP-glucuronic acid and UDP-N-acetylglucosamine precursors. The phosphorylated 121,000-Mr subunit possessed two binding sites with equal affinity towards UDP-glucuronic acid and UDP-N-acetylglucosamine but did not possess UDP-moiety-cleavage activity. The phosphorylation of 200,000-Mr subunit by an intrinsic kinase activity of the protein molecule appeared to elicit its oligosaccharide-synthesizing activity, whereas phosphorylation of 121,000-Mr subunits, presumably carried out in vivo, abolished this activity of the protein molecule. The oligosaccharides synthesized by the protein were about Mr 5000 and about 12 disaccharide units in length. Neither nucleotide sugars nor glycosyl residues nor newly synthesized oligosaccharides were bound covalently to the protein molecule. The UDP moiety of nucleotide sugar precursors did not constitute a link between protein molecule and oligosaccharide during its synthesis. Although isolated 442,000-Mr protein did not synthesize high-Mr hyaluronate in vitro, this protein molecule can be considered as a constituent of membrane-bound hyaluronate synthase complex because of its observed properties.

scholarly journals Analysis of cell-growth-phase-related variations in hyaluronate synthase activity of isolated plasma-membrane fractions of cultured human skin fibroblasts

1986 ◽  
Vol 237 (2) ◽  
pp. 333-342 ◽  
Author(s):  
N Mian
Keyword(s):  
Plasma Membrane ◽  
Growth Phase ◽  
Plasma Membranes ◽  
Glucuronic Acid ◽  
Stationary Growth Phase ◽  
Binding Activity ◽  
Exponential Growth Phase ◽  
Human Skin Fibroblasts ◽  
Stationary Growth ◽  
Hyaluronate Synthase

Hyaluronate synthase activity is localized exclusively in plasma-membrane fractions of cultured human skin fibroblasts. The enzyme activity of plasma membranes prepared from exponential-growth-phase cells was about 6.5 times that of stationary-growth-phase cells. Hyaluronate synthase from exponential-growth-phase cells exhibited lower Km and higher Vmax. values for both UDP-N-acetylglucosamine and UDP-glucuronic acid and higher rate of elongation of hyaluronate chains compared with the enzyme from stationary-growth-phase cells. Hyaluronate synthase exhibited an extremely short half-life, 2.2 h and 3.8 h respectively when cells were treated with cycloheximide and actinomycin D. The cell-growth-phase-dependent variations in hyaluronate synthase activity appear to be due to its high turnover rate as well as due to some post-translational modification of the enzyme protein as cells progress from early exponential to stationary growth phase. The isolated plasma membranes contained a protein (Mr approx. 450,000) that was selectively autophosphorylated from [gamma-32P]ATP in vitro in the presence of hyaluronate precursors in the reaction mixture and that also exhibited some hyaluronate-synthesis-related properties. The 32P-labelled protein isolated from plasma membranes of exponentially growing cells expressed an efficient UDP-[14C]glucuronic acid- and UDP-N-acetyl[3H]glucosamine-binding activity and was able to synthesize oligosaccharides (Mr 5000) of [14C]glucuronic acid and N-acetyl[3H]glucosamine residues. The corresponding protein of stationary-growth-phase cells, which expressed much higher nucleotide-sugar-precursor-binding activity, appeared to have lost its oligosaccharide-synthesizing activity.

Sign in / Sign up

Export Citation Format

Share Document