Purification, Cloning, and Expression of Human Plasma Hyaluronidase

Frost, Gregory I.; Csóka, Tony B.; Wong, Tim; Stern, Robert

doi:10.1006/bbrc.1997.6773

Cited by 221 publications

(134 citation statements)

References 22 publications

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“…Human Hyal-1 hydrolyzes HA in a size-independent manner to tetrasaccharides, whereas hHyal-2 specifically degrades large HA polymers into 20 kDa fragments (44). The hHyal-1 activity optimum is at pH 3.8, hHyal-2 at pH 5.6-7.0 (21,43,44), and the bovine PH-20 pH profile is bimodal, with maxima at 4.5 and 7.5 (21). While the sequences of the reaction centers are conserved in all family members, residues at the peripheries of the active site clefts are more divergent.…”

Section: Discussionmentioning

confidence: 99%

Structure of Human Hyaluronidase-1, a Hyaluronan Hydrolyzing Enzyme Involved in Tumor Growth and Angiogenesis^,

2007

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Mammalian hyaluronidases hydrolyze hyaluronan, a polysaccharide of diverse physiological roles found in all tissues and body fluids. In addition to its function in normal cellular hyaluronan turnover, human hyaluronidase-1 is implicated in cancer proliferation, angiogenesis, and inflammatory diseases; its expression is up-regulated in advanced stages of bladder cancer, whereas the expression of the alternative splice-variants is down-regulated. The crystal structure reveals a molecule composed of two closely associated domains: a catalytic domain that adopts a distorted ( /R) 8 barrel resembling that of bee venom hyaluronidase, and a novel, EGF-like domain, characteristic of involvement in protein-protein interactions and regulatory processes. The structure shows that the fold of this unique EGF-like domain is intact in four alternative splice-variants, whereas the catalytic domain is likely to be unfolded. Thus, these variants may function by competing with the full-length enzyme for the putative protein partner and regulating enzymatic activity in healthy cells.Hyaluronan (HA 1 ) is a linear, unsulfated, negatively charged, glycosaminoglycan formed from ∼2000-25000 repeating disaccharide units of D-glucuronic acid (GlcUA) and N-acetyl-D-glucosamine (GlcNAc) with [GlcUA-( 1f3)-GlcNAc-( 1f4)] n linkages. HA is the major component of cartilage and serves as a joint lubricant. It controls water homeostasis in tissues and the extracellular matrix, which affects cell motility and the distribution and transport of plasma proteins. HA is also implicated in cell proliferation, differentiation, cell-cell recognition, tumor growth and invasion, angiogenesis, and inflammatory responses (1-3). The cellular role of HA and the HA-mediated signal transduction pathways depends on its size (recently reviewed by Stern et al. (4)). Large HA polymers function in organizing the extracellular matrix and serve as lubricant and shock absorber. In shock, septicemia, post surgery, blood loss, and burns, the level of circulating high molecular mass HA increases. These HA polymers are antiangiogenic, immunosuppressive, and anti-inflammatory. In contrast, HA fragments of intermediate sizes are involved in the body's alarm system. They stimulate angiogenesis and inflammatory reactions and facilitate cancer progression and invasion (4). Such HA fragments activate several cytoplasmic and extracellular signal transduction pathways associated with Raf-1 kinase, MAP kinase, and ERK. Short HA oligosaccharides are antiapoptotic and inducers of heat shock proteins. They regulate a different set of signaling molecules than the intermediate size HA including Erb2, PTEN phosphatase, and PI 3 kinase (4).Mammalian hyaluronidases are endo--N-acetyl-hexosaminidases (EC 3.2

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Section: Discussionmentioning

confidence: 99%

Structure of Human Hyaluronidase-1, a Hyaluronan Hydrolyzing Enzyme Involved in Tumor Growth and Angiogenesis^,

2007

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“…PH20 is unique as a membrane-bound, neutral active enzyme with high specific activity toward HA in vitro and in vivo, which is capable of degrading matrix-associated megadalton HA polymers to small tetrasaccharide and hexasaccharide products (16)(17)(18)(19). Given the rapid endogenous turnover of HA, accumulation in tumors could involve disequilibrium between the enzymes that synthesize HA and the enzymes that catabolize HA or receptors for cellular uptake, which subsequently mediate intracellular metabolism (20)(21)(22)(23).…”

Section: Introductionmentioning

confidence: 99%

Enzymatic Depletion of Tumor Hyaluronan Induces Antitumor Responses in Preclinical Animal Models

Thompson

Shepard²,

O’Connor³

et al. 2010

Molecular Cancer Therapeutics

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255

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Hyaluronan (HA) is a glycosaminoglycan polymer that often accumulates in malignancy. Megadalton complexes of HA with proteoglycans create a hydrated connective tissue matrix, which may play an important role in tumor stroma formation. Through its colloid osmotic effects, HA complexes contribute to tumor interstitial fluid pressure, limiting the effect of therapeutic molecules on malignant cells. The therapeutic potential of enzymatic remodeling of the tumor microenvironment through HA depletion was initially investigated using a recombinant human HA-degrading enzyme, rHuPH20, which removed HA-dependent tumor cell extracellular matrices in vitro. However, rHuPH20 showed a short serum half-life (t 1/2 < 3 minutes), making depletion of tumor HA in vivo impractical. A pegylated variant of rHuPH20, PEGPH20, was therefore evaluated. Pegylation improved serum half-life (t 1/2 = 10.3 hours), making it feasible to probe the effects of sustained HA depletion on tumor physiology. In high-HA prostate PC3 tumors, i.v. administration of PEGPH20 depleted tumor HA, decreased tumor interstitial fluid pressure by 84%, decreased water content by 7%, decompressed tumor vessels, and increased tumor vascular area >3-fold. Following repeat PEGPH20 administration, tumor growth was significantly inhibited (tumor growth inhibition, 70%). Furthermore, PEGPH20 enhanced both docetaxel and liposomal doxorubicin activity in PC3 tumors (P < 0.05) but did not significantly improve the activity of docetaxel in low-HA prostate DU145 tumors. The ability of PEGPH20 to enhance chemotherapy efficacy is likely due to increased drug perfusion combined with other tumor structural changes. These results support enzymatic remodeling of the tumor stroma with PEGPH20 to treat tumors characterized by the accumulation of HA. Mol Cancer Ther; 9(11); 3052-64. ©2010 AACR.

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“…Hyal1 is the predominant hyaluronidase in human plasma. 23 Hyal3 and PH-20 are highly expressed in testis and are most likely involved in fertilization. Increased expression of a hyaluronidase similar to that of PH-20 has been reported in metastatic human melanoma, glioblastoma and colon carcinoma, 24 whereas a hyaluronidase similar to Hyal1 was detected in bladder carcinomas.…”

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confidence: 99%

Expression of hyaluronan synthase 2 or hyaluronidase 1 differentially affect the growth rate of transplantable colon carcinoma cell tumors

Jacobson

Rahmanian

Rubin

et al. 2002

Intl Journal of Cancer

112

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Advanced colorectal cancers are often associated with elevated amounts of hyaluronan. To investigate the importance of hyaluronan in colon carcinoma tumor progression, we have expressed by stable transfection hyaluronan synthase 2 (Has2) and hyaluronidase 1 (Hyal1) in the rat colon carcinoma cell line, PROb. We found that hyaluronan overproduction led to a higher growth rate of tumor cells in vitro, and to a faster development of transplantable tumors in syngeneic rats, compared to the mock-transfectants. Has2 transfected PROb cells gave rise to tumors that were significantly less vascularized, but had a significantly larger viable tumor fraction compared to tumors generated from mocktransfectants. In contrast, Hyal1 overexpression suppressed the growth rate of tumor cells both in vitro and in vivo. Moreover, tumors derived from Hyal1-transfected cells had a significantly larger necrotic area than tumors derived from mock-and Has2-transfectants. Our study demonstrates that Has2 overproduction promotes tumorigenicity, whereas Hyal1 overexpression suppresses tumorigenicity in an experimental model for colon carcinoma.

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Purification, Cloning, and Expression of Human Plasma Hyaluronidase

Cited by 221 publications

References 22 publications

Structure of Human Hyaluronidase-1, a Hyaluronan Hydrolyzing Enzyme Involved in Tumor Growth and Angiogenesis^,

Structure of Human Hyaluronidase-1, a Hyaluronan Hydrolyzing Enzyme Involved in Tumor Growth and Angiogenesis^,

Enzymatic Depletion of Tumor Hyaluronan Induces Antitumor Responses in Preclinical Animal Models

Expression of hyaluronan synthase 2 or hyaluronidase 1 differentially affect the growth rate of transplantable colon carcinoma cell tumors

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Purification, Cloning, and Expression of Human Plasma Hyaluronidase

Cited by 221 publications

References 22 publications

Structure of Human Hyaluronidase-1, a Hyaluronan Hydrolyzing Enzyme Involved in Tumor Growth and Angiogenesis,

Structure of Human Hyaluronidase-1, a Hyaluronan Hydrolyzing Enzyme Involved in Tumor Growth and Angiogenesis,

Enzymatic Depletion of Tumor Hyaluronan Induces Antitumor Responses in Preclinical Animal Models

Expression of hyaluronan synthase 2 or hyaluronidase 1 differentially affect the growth rate of transplantable colon carcinoma cell tumors

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Structure of Human Hyaluronidase-1, a Hyaluronan Hydrolyzing Enzyme Involved in Tumor Growth and Angiogenesis^,

Structure of Human Hyaluronidase-1, a Hyaluronan Hydrolyzing Enzyme Involved in Tumor Growth and Angiogenesis^,