1995 Fiscal Year Annual Research Report

ヒト赤血球膜蛋白分画4.2の正常と病態に関する遺伝子分子生化学的、電顕的研究

Research Project

Project/Area Number	07670180
Research Category	Grant-in-Aid for General Scientific Research (C)
Research Institution	Kawasaki Medical School
Principal Investigator	神崎暁郎川崎医科大学, 医学部, 講師 (40148698)
Co-Investigator(Kenkyū-buntansha)	井上孝文川崎医科大学, 医学部, 助手 (60203238) 和田秀穂川崎医科大学, 医学部, 講師 (70191830) 八幡義人川崎医科大学, 医学部, 教授 (70069011)
Keywords	Red cell membrane proteins / Band4.2 / Morphogenesis / Gene mutation / Microelectrongraphy
Research Abstract	ヒト赤芽球レベルにおける赤血球膜蛋白band4.2 (P4.2)の遺伝子と蛋白発現に関し、生化学的、電顕的検討を行うことで、膜骨格蛋白におけるP4.2の生理的意義を明らかにする目的で、研究を遂行した。 (1)P4.2蛋白発現:(a)和田らの赤芽球液体培養法を用い、正常のP4.2発現と赤芽球の分化・成熟とを経時的に検討した。その結果、主要膜蛋白の中でP4.2の発現が特異的に最も遅く、Ortho chromatic erythroblastであることが判明した。(b)正常ヒト骨髄芽球のP4.2発現研究成績からも(a)の結果が裏付けられ、in vivoの赤芽球で立証された。 (二)P4.2蛋白の細胞内局在:電顕的検討により、P4.2の特異的な細胞内局在として、赤芽球が脱核する際、核放出部位もしくは、その周辺の細胞膜に本蛋白が集合することが判明した。これは、P4.2蛋白が脱核する際に重要な作用を司っている可能性を示唆する所見と考えられた。 (3)P4.2遺伝子発現:少なくとも5種のisoformが存在し、今回、exon5のsplice outを受けたisoformがあることが発見された。また、これら遺伝子に対応する蛋白は、wild type (72kD)に加え、63kDと59kDの存在が確認された。 (4)Band4.2異常症の遺伝子解析および病態解析と、band4.2蛋白の構造と機能解析:既に判明しているNippon Typeのband4.2欠損症に加えて、band4.2 Komatsu (523GAT→TAT ; homozygote)、band4.2 Shiga(317CGC→TGC/142GCT→ACT ; compound heterozygote)を同定した。さらに、band4.2の生理的機能に関して、これらのband4.2欠損症の電顕学的、物性学的手法による病態研究を通じて、P4.2が細胞骨格蛋白(特にspectrin)に結合する可能性が高いことが判明した。

Research Products

(11 results)

All Other

All Publications (11 results)

[Publications] Kanzaki, A.: "Band 4.2 Komatsu:523GAT→TAT (175Asp→Tyr) in exon 4 of the band 4.2 gene associated with total deficiency of band 4.2, hemolytic anemia with ovalostomatocytosis and marked disruption of the cytoskeletal network." International Journal of Hematology. 61. 165-178 (1995)
[Publications] Kanzaki, A.: "Band 4.2 Shiga : 317CGC→TGC in compound heterozygotes with 142 GCT→ACT results in band 4.2 deficiency and microspherocytosis." British Journal of Haematology. 91. 333-340 (1995)
[Publications] Marechal, J.: "Ethnic distribution of allele α^<LELY>, alow expression allele of red cell spectrin α-gene." British Journal Haematology. 90. 553-556 (1995)
[Publications] Okamoto, N.: "Hereditary spherocytic anemia with deletion of the short arm of chromosome 8." American Journal of Medical Genetics. 58. 225-229 (1995)
[Publications] Kanzaki, A.: "The definitive role of band 4.2 in completing whole assembly of red cell membrane structure by the latest expression with five isoforms among membrane proteins at the terminal erythroid maturation." Blood. 86. 468a (1995)
[Publications] Yawata, Y.: "Complete band 3 deficiency in cattle : A model for hereditary spherocytosis with striking instability of cytoskeletal metwork with marked exo-and endocytosis" Blood. 86. 468a (1995)
[Publications] Yawata, Y.: "Electron microscopic evidence of impaired intramembrane particles and of instability of cytoskeletal network in band 4.2 deficiency in human red cells." Cell Motility and the Cytoskeleton. 33. 95-105 (1996)
[Publications] Maillet, P.: "A stop codon in exon 30 of β-spectrin gene associated with hereditary elliptocytosis in spectrin Nagoya." Human Mutation. (in press). (1996)
[Publications] Kanzaki, A.: "The completion of whole assembly of red cell membrane structure by the latest expression of band 4.2 with five isoforms among membrane proteins at the terminal erythroid maturation." Proceedings of the International Symposium on Membrane Proteins : Structure, Function and Expression Control (Feb. 7-9, 1996, Fukuoka). 88 (1996)
[Publications] Inaba, M.: "Defective anion transport and marked spherocytosis with membrane instability caused by hereditary total deficiency of red cell band 3 due to a nonsense mutation." Journal of Clinical Investigation. (in press). (1996)
[Publications] Yawata, Y.: "Novel evidence of binding of red cell membrane protein band 4.2 to spectrins critical for the final membrane assembly and membrane stability." Proceedings of the International Symposium on Membrane Proteins : Structure, Function and Expression Control (Feb. 7-9, 1996, Fukuoka). 87 (1996)

1995 Fiscal Year Annual Research Report

ヒト赤血球膜蛋白分画4.2の正常と病態に関する遺伝子分子生化学的、電顕的研究

Principal Investigator

神崎 暁郎 川崎医科大学, 医学部, 講師 (40148698)

Research Products

[Publications] Kanzaki, A.: "Band 4.2 Shiga : 317CGC→TGC in compound heterozygotes with 142 GCT→ACT results in band 4.2 deficiency and microspherocytosis." British Journal of Haematology. 91. 333-340 (1995)

[Publications] Marechal, J.: "Ethnic distribution of allele α^<LELY>, alow expression allele of red cell spectrin α-gene." British Journal Haematology. 90. 553-556 (1995)

[Publications] Okamoto, N.: "Hereditary spherocytic anemia with deletion of the short arm of chromosome 8." American Journal of Medical Genetics. 58. 225-229 (1995)

[Publications] Kanzaki, A.: "The definitive role of band 4.2 in completing whole assembly of red cell membrane structure by the latest expression with five isoforms among membrane proteins at the terminal erythroid maturation." Blood. 86. 468a (1995)

[Publications] Yawata, Y.: "Complete band 3 deficiency in cattle : A model for hereditary spherocytosis with striking instability of cytoskeletal metwork with marked exo-and endocytosis" Blood. 86. 468a (1995)

[Publications] Yawata, Y.: "Electron microscopic evidence of impaired intramembrane particles and of instability of cytoskeletal network in band 4.2 deficiency in human red cells." Cell Motility and the Cytoskeleton. 33. 95-105 (1996)

[Publications] Maillet, P.: "A stop codon in exon 30 of β-spectrin gene associated with hereditary elliptocytosis in spectrin Nagoya." Human Mutation. (in press). (1996)

[Publications] Inaba, M.: "Defective anion transport and marked spherocytosis with membrane instability caused by hereditary total deficiency of red cell band 3 due to a nonsense mutation." Journal of Clinical Investigation. (in press). (1996)

神崎暁郎川崎医科大学, 医学部, 講師 (40148698)