Purification and properties of an antivenom factor from the plasma of the South American rattlesnake (Crotalus durissus terrificus)

Fortes-Dias, Consuelo Latorre; Fonseca, B.C.B.; Kochva, Elazar; Diniz, Carlos R.

doi:10.1016/0041-0101(91)90082-3

Cited by 58 publications

(28 citation statements)

References 32 publications

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“…NAI is the only PLI to inhibit the enzymatic activity of the hsPLA 2 -II enzyme. Crotalus neutralizing factor, purified from Crotalus durissus terrificus (South American rattlesnake) serum (32)(33)(34), was tested against porcine pancreatic PLA 2 (group I), and Trimeresurus flavoviridis PLI, purified from T. flavoviridis (Habu) serum (37) was tested on bovine pancreatic PLA 2 ; both PLIs were incapable of inhibiting enzymatic activity (33,37).…”

Section: Discussionmentioning

confidence: 99%

Functional Characteristics of a Phospholipase A2Inhibitor from Notechis ater Serum

Hains

Sung

Tseng

et al. 2000

Journal of Biological Chemistry

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A phospholipase A 2 inhibitor has been purified from the serum of Notechis ater using DEAE-Sephacel chromatography. The inhibitor was found to be composed of two protein subunits (␣ and ␤) that form the intact complex of approximately 110 kDa. The ␣-chain is a 30-kDa glycoprotein and the ␤-chain a nonglycosylated, 25-kDa protein. N-terminal sequence analysis reveals a high level of homology to other snake phospholipase A 2 inhibitors. The inhibitor was shown to be extremely pH and temperature stable. The inhibitor was tested against a wide variety of phospholipase A 2 enzymes and inhibited the enzymatic activity of all phospholipase A 2 enzymes tested, binding with micromole to nanomole affinity. Furthermore, the inhibitor was compared with the Eli-Lilly compound LY311727 and found to have a higher affinity for human secretory nonpancreatic phospholipase A 2 than this chemical inhibitor. The role of the carbohydrate moiety was investigated and found not to affect the in vitro function of the inhibitor.For many years research on PLA 2 1 enzymes has centred on snake venom PLA 2 s, largely because of the fact that PLA 2 enzymes are extremely abundant in snake venoms and are easily purified. PLA 2 enzymes have also been identified and purified from bovine, porcine, and human pancreas (1-3) and in human synovial fluid aspirates from rheumatoid and osteoarthritis patients (4, 5). PLA 2 enzymes have been grouped according to structural and sequence characteristics. Pancreatic PLA 2 enzymes belong to group I, whereas the human secretory nonpancreatic PLA 2 is a group II enzyme. PLA 2 enzymes from snake venom belong to group I or II (6 -8).Recently PLA 2 enzymes have been implicated as playing a role in a range of diseases including rheumatoid and osteoarthritis, asthma, acute pancreatitis, psoriasis, multiple organ failure, septic shock, and adult respiratory distress syndrome (9 -11). Several review articles (9,10,(12)(13)(14) have examined the physiological and potential disease role of hsPLA 2 -II. Of major importance are the reaction by-products of PLA 2 activity, a free fatty acid and a lysophospholipid. The fatty acid is often arachidonic acid, one of the main constituents of the cell membrane in several tissues (15)(16)(17). Arachidonic acid is the precursor for inflammatory mediators such as thromboxanes, leukotrienes, and prostaglandins (18,19). Lysophospholipids are the precursor of platelet activating factor (20).The suggestion that PLA 2 enzymes play a role in diseases process has intensified research on PLA 2 inhibitors. Many investigators have purified and characterized PLIs from numerous sources including plant, fungi, and bacteria (21-28). A large research effort has gone into the design of synthetic compounds, such as the Eli-Lilly compound LY311727, which specifically inhibits hsPLA 2 -II (29). Additionally, a number of PLIs have been purified from the serum of elapid and crotalid snakes. These PLIs have all been shown to be homologous or heterologous complexes that interact with PLA 2 enzymes to inhib...

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Section: Discussionmentioning

confidence: 99%

Functional Characteristics of a Phospholipase A2Inhibitor from Notechis ater Serum

Hains

Sung

Tseng

et al. 2000

Journal of Biological Chemistry

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“…The molecular mass was calculated to be 20250 Da from the cDNA sequence. Although a putative polyadenylation site has not been found in CNF cDNA [18], there is a putative polyadenylation signal in PLI-I cDNA (Fig. 6).…”

Section: Inhibitory Effects Of the Binding Proteins On Their Target Pmentioning

confidence: 99%

Characterization and Evolution of a Gene Encoding a Trimeresums flavoviridis Serum Protein that Inhibits Basic Phospholipase A₂ Isozymes in the Snake's Venom

Nobuhisa

Inamasu

Nakai

et al. 1997

European Journal of Biochemistry

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The proteins that bind phospholipase A2 (PLA2) isozymes of Trimeresurus flavoviridis (habu snake, crotalinae) venom were ractionated from sera on four columns, each conjugated with one of four PLA2 isozymes. Five proteins, termed PLA2 inhibitors (PLI) I–V, were obtained as the binding components. The combinations of the binding components differed depending on the PLA2 isozymes. PLI‐IV and PLI‐V correspond to PLI‐A and PLI‐B, respectively, which were known to bind to a major [Asp49]PLA2, PLA2, and contained a segment similar to the carbohydrate‐recognition domain of C‐type lectins. PLI‐I, which is a major component of inhibitory proteins against three basic PLA2 isozymes, PLA‐B (a basic [Asp49]PLA2) and basic proteins I and II (both [Lys49]PLA,s), has been isolated, and its partial amino acid sequence has been determined. A cDNA encoding PLI‐I was isolated from a T. flavoviridis liver cDNA library and sequenced. PLI‐I cDNA encoded 200 amino acid residues, including a signal peptide of 19 amino acid residues. One sugar chain was predicted to occur at position 157. A gene coding for PLI‐I was isolated. It is 9.6‐kb long and consists of five exons and four introns. Comparison of the exonintron structure of the PLI‐I gene with those of genes encoding urokinaseype‐plasminogen‐activator receptor (uPAR), Ly‐6, CD59 and neurotoxins showed that they have characteristic unit encoding approximately 90 amino acid residues, which is divided over two exons. This strongly suggests that the PLI‐I gene belongs to the uPAR, Ly‐6, CD59 and neurotoxin gene family. There are two types of structurally different inhibitors against PLA2 isozymes in T. flavoviridis serum with different evolutionary origins.

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“…These PLIs have been purified from Naja naja kaouthia (Thailand cobra [23,24]), Laticauda semifasciata (Sea Krait [25]), Crotalus durissus terrificus (South American rattlesnake [26][27][28]), Trimeresurus fla o iridis (Habu [29][30][31][32]), Agkistrodon blomhoffii siniticus (Chinese mamushi [33,34]) and Bothrops asper (fer-delance [35]). Additionally, an inhibitory serum factor from Vipera palaestinae (Palaestinian viper) that shares many of the characteristics of PLIs has been identified [36] ; however, it has not been clearly shown that this protein inhibits PLA # enzymes.…”

Section: Introductionmentioning

confidence: 99%

Purification and inhibitory profile of phospholipase A2 inhibitors from Australian elapid sera

Hains

Broady

2000

Biochemical Journal

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Although the resistance of snakes to their own venom is well known, until now no investigators have examined the serum of Australian snakes. Here we describe the identification and purification of a range of phospholipase A2 (PLA2) inhibitors from the serum of Australian elapids. All PLA2 inhibitors were composed of two protein chains, an α-chain and a β-chain. The α-chains were approx. 22.5 kDa in size and variably glycosylated, whereas the β-chains were approx. 19.8 kDa in size and not glycosylated. Identification of isoforms of the two subunit chains was significant because three of the six sera examined were from single snake specimens. In addition, the glycosylation patterns of the α-chains were thoroughly investigated in these unpooled sera. The functional and structural properties of the purified inhibitors were studied. Uniquely, a snake PLA2 inhibitor was found to inhibit human type II PLA2 enzyme, which has implications for the treatment of the many diseases in which PLA2 enzymes have been implicated. Further, we demonstrate that the inhibitor forms a non-covalent association with a purified PLA2 enzyme. Finally, the purified PLA2 inhibitor was shown to protect in vivo against the lethal affects of a homologous PLA2 enzyme, suggesting a role for PLA2 inhibitors in the treatment of snake bite victims.

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Purification and properties of an antivenom factor from the plasma of the South American rattlesnake (Crotalus durissus terrificus)

Cited by 58 publications

References 32 publications

Functional Characteristics of a Phospholipase A2Inhibitor from Notechis ater Serum

Functional Characteristics of a Phospholipase A2Inhibitor from Notechis ater Serum

Characterization and Evolution of a Gene Encoding a Trimeresums flavoviridis Serum Protein that Inhibits Basic Phospholipase A₂ Isozymes in the Snake's Venom

Purification and inhibitory profile of phospholipase A2 inhibitors from Australian elapid sera

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Purification and properties of an antivenom factor from the plasma of the South American rattlesnake (Crotalus durissus terrificus)

Cited by 58 publications

References 32 publications

Functional Characteristics of a Phospholipase A2Inhibitor from Notechis ater Serum

Functional Characteristics of a Phospholipase A2Inhibitor from Notechis ater Serum

Characterization and Evolution of a Gene Encoding a Trimeresums flavoviridis Serum Protein that Inhibits Basic Phospholipase A2 Isozymes in the Snake's Venom

Purification and inhibitory profile of phospholipase A2 inhibitors from Australian elapid sera

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Characterization and Evolution of a Gene Encoding a Trimeresums flavoviridis Serum Protein that Inhibits Basic Phospholipase A₂ Isozymes in the Snake's Venom