Purification of streptokinase by affinity chromatography using human plasminogen = 친화성 크로마토그래피를 이용한 스트렙토키나제의 분리

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A method for preparing a respectable quality streptokinase in a convenient manner was described. Streptokinase is important for enzyme theraphy, because it lyses blood clots by activating plasminogen to the fibrinolytic enzyme, plasmin. For the crystallography to determine its tertiary structure, the preparation of pure streptokinase is needed. Human plasminogen was purified from Cohn fraction III by the affinity chromatography with Llysine-substituted Sepharose CL-4B. This plasminogen was used as a ligand for streptokinase affinity chromatography. PBS (pH7.4) and Glycine (pH2.5) were used as washing and elution buffer, respectively. To obtain a single peak, $\varepsilon$-amino-N-caproic acid ($\varepsilon$-ACA) was added to both washing and elution buffers. The function of $\varepsilon$-ACA is the prevention of streptokinase fragmentation during the plasminogen activation. To study the role of Gly 24 in streptokinase, mutant streptokinaess, His 24, Ala 24 and Glu 24, were purified and tested for specific activity and protease activity with various artificial substrates. The specific activity of Ala-SK was about 76.6\% of Gly-SK, and His-SK and Glu-SK lost nearly all activator activity. And any protease activity against substrates tested so far has not been detected in wild-type and mutant streptokinases.
Advisors
Byun, Si-Myung변시명
Description
한국과학기술원 : 생명과학과,
Publisher
한국과학기술원
Issue Date
1993
Identifier
68308/325007 / 000911135
Language
eng
Description

학위논문(석사) - 한국과학기술원 : 생명과학과, 1993.2, [ vii, 48 p. ]

Keywords

단백질 분리.

URI
http://hdl.handle.net/10203/28396
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=68308&flag=dissertation
Appears in Collection
BS-Theses_Master(석사논문)
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