PURIFICATION OF CYSTEINE-RICH BIOACTIVE PEPTIDES FROM LEUKOCYTES BY CONTINUOUS ACID-UREA POLYACRYLAMIDE-GEL ELECTROPHORESIS

被引：59

作者：

HARWIG, SSL ^{[1
]}

CHEN, NP ^{[1
]}

PARK, ASK ^{[1
]}

LEHRER, RI ^{[1
]}

机构：

[1] UNIV CALIF LOS ANGELES, WADSWORTH VET ADM HOSP, DEPT MED, LOS ANGELES, CA 90025 USA

来源：

ANALYTICAL BIOCHEMISTRY | 1993年 / 208卷 / 02期

关键词：

D O I：

10.1006/abio.1993.1065

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A new continuous acid-urea-polyacrylamide gel electrophoresis (CAU-PAGE) preparative method was developed and used to purify rabbit and human defensins. With it, we identified two post-translationally modified forms of rabbit defensins NP-1 and NP-2, and purified a processed RANTES (β-intercrine) peptide from leukophoresed human leukocytes. CAU-PAGE afforded approximately 70% recovery of rabbit defensin NP-5. The recovered defensins were not N-terminally modified, and their in vitro antimicrobial activity was equivalent to that of defensins purified by previously described chromatographic methods. Since CAU-PAGE is performed under nonreducing conditions, it should be especially useful for purifying cationic peptides with intramolecular disulfide bonds, such as defensins and α or β-intercrines. © 1994 Academic Press, Inc. All rights reserved.

引用

页码：382 / 386

页数：5

共 22 条

[1] GRANULINS, A NOVEL CLASS OF PEPTIDE FROM LEUKOCYTES [J].

BATEMAN, A ;

BELCOURT, D ;

BENNETT, H ;

LAZURE, C ;

SOLOMON, S .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1990, 173 (03) :1161-1168

[2] RAPID ANALYSIS OF AMINO-ACIDS USING PRE-COLUMN DERIVATIZATION [J].

BIDLINGMEYER, BA ;

COHEN, SA ;

TARVIN, TL .

JOURNAL OF CHROMATOGRAPHY, 1984, 336 (01) :93-104

[3] DEFENSINS - NATURAL PEPTIDE ANTIBIOTICS OF HUMAN-NEUTROPHILS [J].

GANZ, T ;

SELSTED, ME ;

SZKLAREK, D ;

HARWIG, SSL ;

DAHER, K ;

BAINTON, DF ;

LEHRER, RI .

JOURNAL OF CLINICAL INVESTIGATION, 1985, 76 (04) :1427-1435

[4]

HAGER KM, 1991, BIO RAD US EG B, V1685

[5]

HARWIG SSL, 1992, BLOOD, V79, P1532

[6]

HEINRIKSON RL, 1984, ANAL BIOCHEM, V136, P65, DOI 10.1016/0003-2697(84)90307-5

[7] INSECT IMMUNITY - ISOLATION FROM IMMUNE BLOOD OF THE DIPTERAN PHORMIA-TERRANOVAE OF 2 INSECT ANTIBACTERIAL PEPTIDES WITH SEQUENCE HOMOLOGY TO RABBIT LUNG MACROPHAGE BACTERICIDAL PEPTIDES [J].

LAMBERT, J ;

KEPPI, E ;

DIMARCQ, JL ;

WICKER, C ;

REICHHART, JM ;

DUNBAR, B ;

LEPAGE, P ;

VANDORSSELAER, A ;

HOFFMANN, J ;

FOTHERGILL, J ;

HOFFMANN, D .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (01) :262-266

[8] ULTRASENSITIVE ASSAYS FOR ENDOGENOUS ANTIMICROBIAL POLYPEPTIDES [J].

LEHRER, RI ;

ROSENMAN, M ;

HARWIG, SSSL ;

JACKSON, R ;

EISENHAUER, P .

JOURNAL OF IMMUNOLOGICAL METHODS, 1991, 137 (02) :167-173

[9] DEFENSINS - ENDOGENOUS ANTIBIOTIC PEPTIDES OF ANIMAL-CELLS [J].

LEHRER, RI ;

GANZ, T ;

SELSTED, ME .

CELL, 1991, 64 (02) :229-230

[10]

MATSUDAIRA P, 1987, J BIOL CHEM, V262, P10035

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