Elegantin and albolabrin purified peptides from viper venoms; homologies with the RGDS domain of fibrinogen and von Willebrand factor

Williams, Janice A.; Rucinski, Boguslaw; Holt, John Clifford; Niewiarowski, Stefan

doi:10.1016/0167-4838(90)90229-9

Cited by 85 publications

(61 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…1). This pattern ~tends from the first to the last cysteine in the pcptide, and leaves only a few N-terminal (one to five:) and C-terminal (six to ten) amino acids beyond these loops [1][2][3][4][5][6][7][8][9][10]. In all thr~ disintogrins, two disulfide bridges form an interesting motif with respect to the RGD sequence.…”

Section: Discussionmentioning

confidence: 99%

The disulfide bridge pattern of snake venom disintegrins, flavoridin and echistatin

et al. 1992

Self Cite

View full text Add to dashboard Cite

Flavoridin and echistatin, isolated from the-venom of Trh,¢resuruxflararlridis and Echix ¢¢zrh~utu$. respectively. I~lonB ¢o tilt disintel;rin family of inttgrin p~ and 0~ inhibitors ol" low molecular w¢isht gGD.¢ontainin8, cy,qeinc.vith poptid=s. Sin= disulfide bonds ar¢ criti~l for expression ot" biological activity, we sought to determine their location in these two proteins. In fluvoridin, direct cvidcn=: for the exist¢n¢¢ o1. linkug~ between Cys'-Cy= =" and betwcgn Cys ° and Cys" was obtained by analysis of proteolyti¢ products, and indirect evidence sullg¢sts links between Cys'-Cys =+ and Cys~>-Cys ~. In e=histatin, links between Cys'-Cys >~ and Cys'~-Cys ~ were identified by dir=t chemical analysis, 2, MATERIALS AND METHODSSynthetic ethistutin was purchased from Uach=m Inc. (USA) and was used without further purification, Natural echistatin and fla. either natural or synthetic, and flavoridin, (2 rag/m| in lO0 mM NH.,HCO;~. pH 8,0) wcr¢ di;~|cd with TPCK.trypsin (Sigma) at an ¢nxyme:substrato ratio of I:50 (w/w) for 4 h at 3"PC, Dil~tion ol'echistatin was stopl~.d with formic acid (final ¢omazntra. tion 30~ (v/v)), and the styptic popfidcs were scparut¢ by rever~e-phar, e HPLC on a Lichrosphcr JOe RP-l~ (S#M particle size) (Merck, Darmstadt) column cquilibrate.d in a mixture of O.Ig'a (v/v) TFA in wuttr (solution A) and in u=tonilrile (solution B) (95~ A/:~% I~) and ¢luted at I mVmin, at ~rst isocratically for 5 rain and then followed by a linear gradient up to 40'~ B in 70 rain, Th= enzyme in the trypti¢ digest o1" flavoridin was h=at-inactivatcd (1,00"C t'or 5 rain), endnproteinase Asp-N (Boehrin~r.Mannhcim, Indianapolis. IN) was ;tddcd up to u final tnxyme:substra|c ratio o1" h25 (w/w) and incubated 6 h at 37'C, Thereafter, the resuhinll pep tides were separated ;is above. Elation conditions were: 5 rain isoeratirally (100% A) followed by a linear gradient up to 30~ B in 90 rain.The isolated peptid¢s wore chaructedzc.d by amino acid analysis (after acid hydrolysis with 6 N HCI foe 24 h at 110"C using a Biotronic (amino acids aualyzcr) and amino-tem~inal sequence analysis (using an Applied Biosystcms. Sun Francisco. CA) gas-phase scquenc=r mod~l 473A, Mass spectra were recorded with a mats spectrometer MAT 900 (Finni~n MAT. Bremen. FRO) equipped with liquid s¢c-oudaw ion ionization system. RESULTS 3,1. Disu~de bridge pattern of fiavorldinProteolytic digestion of flavoridin with trypsin and endoproteinuse Asp.N gave nine lt'actions (Table I), 316 Published by Efscvicr S¢ietw¢ Publldtcrs B. V,

show abstract

Section: Discussionmentioning

confidence: 99%

The disulfide bridge pattern of snake venom disintegrins, flavoridin and echistatin

et al. 1992

Self Cite

View full text Add to dashboard Cite

show abstract

“…1. The concentrations of eristostatin, echistatin and echistatin D27W that inhibited ADP-induced platelet aggregation (IC~0) determined as previously described [19] were 59+22 nM, 139+17 nM and 83+16 nM, respectively. Synthetic echistatin was identical in this functional assay to native echistatin.…”

Section: Preparation and Characterization Of Disintegrinsmentioning

confidence: 99%

“…Eristostatin and echistatin were purified into homogeneity from the crude venom of Eristocophis macmahoni and Echis carinatus, respectively, using two-step C-18 reversed-phase HPLC by the method of Williams et al [19]. Synthetic echistatinwt and echistatin D27W were provided by Dr. Victor Garsky (Merck Sharp and Dohme Research Laboratories, West Point, PA).…”

Section: Preparation and Characterization Of Disintegrinsmentioning

confidence: 99%

Importance of the structure of the RGD‐containing loop in the disintegrins echistatin and eristostatin for recognition of αIIbβ3 and αvβ3 integrins

et al. 1996

Self Cite

View full text Add to dashboard Cite

show abstract

“…Platelet Aggregation-The concentration of each recombinant or native disintegrin that caused inhibition of platelet aggregation induced by 20 M ADP was determined as described previously (29).…”

Section: Materials-lyophilizedmentioning

confidence: 99%

Structural Requirements of Echistatin for the Recognition of αvβ3 and α5β1Integrins

Wierzbicka-Patynowski¹,

Niewiarowski²,

Marcinkiewicz³

et al. 1999

Journal of Biological Chemistry

Self Cite

135

123

View full text Add to dashboard Cite

There are key differences between the amino acid residues of the RGD loops and the C termini of echistatin, a potent antagonist of ␣ IIb ␤ 3 , ␣ v ␤ 3 and ␣ 5 ␤ 1 , and eristostatin, a similar disintegrin selectively inhibiting ␣ IIb ␤ 3 . In order to identify echistatin motifs required for selective recognition of ␣ v ␤ 3 and ␣ 5 ␤ 1 integrins, we expressed recombinant echistatin, eristostatin, and 15 hybrid molecules. We tested them for their ability to inhibit adhesion of different cell lines to fibronectin and von Willebrand factor and to express ligand-induced binding site epitope. The integrins are a family of cell surface glycoproteins that act as receptors for extracellular matrix (ECM) 1 proteins, or for membrane-bound counter-receptors on other cells. Each integrin is a heterodimer that contains an ␣ and a ␤ subunit, the pairing of which specifies the ligand binding abilities of integrins (1). Integrins can bind adhesive ligands and upon this binding undergo conformational changes leading to the exposure of neoepitopes recognized by specific monoclonal antibodies called anti-ligand-induced binding site (LIBS) antibodies (2).The Arg-Gly-Asp (RGD) sequence is the cell attachment site of a large number of adhesive ECM, blood, and cell surface proteins, and nearly half of the over 20 known integrins recognize this sequence in their adhesion protein ligands (3). In addition to the RGD motif, many adhesive receptors recognize other integrin-binding domains, such as the KQAGDV sequence (4) of fibrinogen ␥-chain, ILDV sequence of the CS1 region of fibronectin (5, 6), and RRETAWA sequence identified from a phage display library (7). It is known that the Arg and Asp residues are necessary but not sufficient to ensure binding activity. The RGD sequence is generally found to be associated with a series of probable ␤-bends, which result in a highly ordered structure. This highly ordered conformation might form the basis of the specific binding of many proteins containing this cell surface recognition sequence. Additional determinants of integrin specificity and the high affinity of RGDcontaining adhesive proteins for integrins may be the result of the specific conformation of the RGD sequence or by the amino acids immediately adjacent to the RGD site, creating an extended RGD locus.Disintegrins are snake venom proteins capable of binding to integrins and interfering with integrin function (8 -10). Disintegrins typically have an RGD sequence as their active site, except for barbourin containing a KGD sequence (11), and a new class of heterodimeric disintegrins such as EC3 and EMF10, containing MLD, VGD, and other recognition motifs (12). It appears that low molecular weight disintegrins, binding to integrins with an affinity approaching that of monoclonal antibodies, may represent a useful probe to identify functionally important motifs in the cell adhesion receptors. Scarborough et al. (13) postulated that the amino acid residue Cterminal to the RGD sequence determines selectivity of disintegrin for an integrin re...

show abstract

Elegantin and albolabrin purified peptides from viper venoms; homologies with the RGDS domain of fibrinogen and von Willebrand factor

Cited by 85 publications

References 27 publications

The disulfide bridge pattern of snake venom disintegrins, flavoridin and echistatin

The disulfide bridge pattern of snake venom disintegrins, flavoridin and echistatin

Importance of the structure of the RGD‐containing loop in the disintegrins echistatin and eristostatin for recognition of αIIbβ3 and αvβ3 integrins

Structural Requirements of Echistatin for the Recognition of αvβ3 and α5β1Integrins

Contact Info

Product

Resources

About