title: Comparison of biological activities of human antithrombins with high-mannose or complex-type nonfucosylated N-linked oligosaccharides.
abstract: The structure of the N-linked oligosaccharides attached to antithrombin (AT) has been shown to affect its anticoagulant activity and pharmacokinetics. Human AT has biantennary complex-type oligosaccharides with the unique feature of lacking a core fucose, which affects its biological activities by changing its heparin-binding affinity. In human plasma, AT circulates as a mixture of the α-form bearing four oligosaccharides and the β-form lacking an oligosaccharide at Asn135. However, it remains unclear how the immature high-mannose-type oligosaccharides produced by mammalian cells affect biological activities of AT. Here, we succeeded in directly comparing the activities between the high-mannose and complex types. Interestingly, although there were no substantial differences in thrombin inhibitory activity, the high-mannose type showed higher heparin-binding affinity. The anticoagulant activities were increased by heparin and correlated with the heparin-binding affinity, resulting in the strongest anticoagulant activity being displayed in the β-form with the high-mannose type. In pharmacokinetic profiling, the high-mannose type showed a much shorter plasma half-life than the complex type. The β-form was found to have a prolonged plasma half-life compared with the α-form for the high-mannose type; conversely, the α-form showed a longer half-life than the β-form for the complex-type. The present study highlights that AT physiological activities are strictly controlled not only by a core fucose at the reducing end but also by the high-mannose-type structures at the nonreducing end. The β-form with the immature high-mannose type appears to function as a more potent anticoagulant than the AT typically found in human plasma, once it emerges in the blood.
authors.affiliation: Bio Process Research and Development Laboratories, Production Division, Kyowa Hakko Kirin Co., Ltd., Takasaki-shi, Gunma 370-0013, Japan.
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Yamada
authors.affiliation: Fuji Research Park, R&D Division, Kyowa Hakko Kirin Co., Ltd., Suntou-gun, Shizuoka 411-8731, Japan.
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Kanda
authors.affiliation: Fuji Research Park, R&D Division, Kyowa Hakko Kirin Co., Ltd., Suntou-gun, Shizuoka 411-8731, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Takayama
authors.affiliation: Fuji Research Park, R&D Division, Kyowa Hakko Kirin Co., Ltd., Suntou-gun, Shizuoka 411-8731, Japan.
authors.forename: Akitoshi
authors.initials: A
authors.lastname: Hashimoto
authors.affiliation: Bio Process Research and Development Laboratories, Production Division, Kyowa Hakko Kirin Co., Ltd., Takasaki-shi, Gunma 370-0013, Japan.
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Sugihara
authors.affiliation: Tokyo Research Park, R&D Division, Kyowa Hakko Kirin Co., Ltd., Machida-shi, Tokyo 194-8533, Japan.
authors.forename: Ai
authors.initials: A
authors.lastname: Satoh-Kubota
authors.affiliation: Tokyo Research Park, R&D Division, Kyowa Hakko Kirin Co., Ltd., Machida-shi, Tokyo 194-8533, Japan.
authors.forename: Eri
authors.initials: E
authors.lastname: Suzuki-Takanami
authors.affiliation: R&D Planning Department, R&D Division.
authors.forename: Keiichi
authors.initials: K
authors.lastname: Yano
authors.affiliation: Tokyo Research Park, R&D Division, Kyowa Hakko Kirin Co., Ltd., Machida-shi, Tokyo 194-8533, Japan shigeru.iida@kyowa-kirin.co.jp.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Iida
authors.affiliation: Immunology & Allergy R&D Unit, R&D Division, Kyowa Hakko Kirin Co., Ltd., Chiyoda-ku, Tokyo 100-8185, Japan.
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Satoh
mesh_terms: D058833:Antithrombin Proteins / Q000737:chemistry / Q000378:metabolism*; D006031:Glycosylation; D006493:Heparin / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008358:Mannose / Q000737:chemistry / Q000378:metabolism*; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D058833:Antithrombin Proteins; D009844:Oligosaccharides; D006493:Heparin; D008358:Mannose
keywords: anticoagulant activities; antithrombin; heparin-binding affinity; high-mannose-type N-linked oligosaccharides; pharmacokinetics