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H M Verheij

Publications and source records attributed to H M Verheij.

125 records · Page 7Linked to original sources
PubMed1978

The primary structure of bovine pancreatic phospholipase A2.

The complete amino acid sequence of bovine phospholipase A2 (EC 3.1.1.4) was determined. This enzyme has a molecular weight of 13 782 and consists of a single polypeptide chain of 123 amino acids cross-linked by seven disulfide bridges. The main fragmentation of the polypeptide chain was accomplished by digesting the reduced and thialaminated derivative of the protein with trypsin, staphylococcal protease and cyanogen bromide. A number of chymotryptic peptides were used for alignment and to obtain overlaps of at least two residues. The sequence of the peptides was determined by Edman degradation by means of direct phenylthiohydantoin identification in combination with identification as dansyl amino acids. Although 71% of all residues of phospholipase A2 from bovine, porcine and equine sources are conserved, bovine phospholipase A2 differs from the others by the total number of residues and by substitutions at 20 (porcine) and 33 (equine) positions.

Amino Acid Sequence↗
PubMed1977

The primary structure of phospholipase A2 from porcine pancreas. A reinvestigation.

The primary structure of porcine pancreatic phospholipase A2 (EC 3.1.1.4) has been reinvestigated. A number of modifications have been introduced including the addition of a 7th disulfide bridge. The structure which is presented here shows a high degree of homology with the amino acid sequence of snake venom and horse pancreas phospholipase A2.

Amino Acid Sequence↗
PubMed1977

Isolation and properties of prophospholipase A2 and phospholipase A2 from horse pancreas and horse pancreatic juice.

Two phospholipases A2 (EC 3.1.1.4) with different isoelectric points have been isolated from horse pancreas in high yield (880 mg/kg tissue). From pancreatic juice the more acidic species was isolated as the sole phospholipase A2. Upon tryptic activation the zymogens release a hepta- and pentapeptide, respectively from the N-terminal part of the protein giving rise to the formation of one single enzyme with a specific activity higher than that of pancreatic phospholipases A2 from other mammalian species. Horse phospholipase A2 differs from the porcine and bovine enzymes with respect to amino acid composition and kinetic properties. The sequence of the first 41 amino acid residues at the N-terminus has been determined by automatic Edman degradation.

Amino Acid Sequence↗
PubMed1977

Amino acid sequence of phospholipase A2 from horse pancreas.

The complete amino acid sequence of phosphlipase A2 (EC 3.1.1.4) from horse pancreas was determined. The protein controls of a single polypeptide chain of 125 amino acids and has a molecular weight of 13,927. The chain is crosslinked by seven disulfide bridges. The sequence was determined by automated Edman degradation of the intact protein and several of the large peptide fragments. Smaller peptides were analyzed by manual Edman degradation. Fragmentation of the peptide chain was accomplished by enzymatic digestion with trypsin, chymotrypsin, and thermolysin. The final overlap was found by digestion of the polypeptide with a staphylococcal protease specific for glutamoyl bonds. Phospholipase A2 from horse pancreas shows homology to snake venom phospholipases A2 and to the enzyme from porcine pancreas, provided that the published amino acid sequence of the porcine phospholipase A2 is revised to some extent.

Amino Acid Sequence↗
PubMed1976

Application of enantiomeric 2-sn-phosphatidylcholines in interfacial enzyme kinetics of lipolysis.

Two enantiomeric 2-sn-phosphatidylcholines containing hexanoyl and dodecanoyl acyl chains have been synthesized, enabling the study of the action of phospholipase A2 (EC 3.1.1.4) at lipid-water interfaces characterized by identical physico-chemical properties. Monolayer kinetics and bulk kinetics in the presence of Triton X-100 micelles were studied but the interpretation of the results is impeded by the fact that interfacial saturation conditions cannot be reached. In contrast, the use of the substrate analog n-tetradecylphosphorylcholine allows the determination of the interfacial kinetic parameters kcat and K*m. Dodecanoic acid is released from the most susceptible isomer about 13 times more rapidly than hexanoic acid from the stereoisomer in spite of the higher K*m of the former. The results are discussed in terms of the particular active site architecture and the possible influence of the "quality of the interface" on the kinetic parameters.

Binding Sites↗
PubMed1972

The structure of a glycerylphosphoryldiglucosyl diglyceride from the lipids of Acholeplasma laidlawii strain B.

1. The phosphatidylglucose structure proposed previously (Smith & Henrikson, 1965) for the glucose-containing phospholipid from Acholeplasma laidlawii is incorrect. 2. The structure now proposed is 3-(sn-glycerol-3-phosphoryl-6'-[O-alpha-d-glucopyranosyl-(1-->2)-O-alpha-d-glucopyranosyl])- sn-1,2-diglyceride, a new type of bacterial lipid. 3. Deacylation of the lipid gave a single water-soluble phosphate ester which could be distinguished on chromatography from synthetic samples of glucosylphosphorylglycerols. 4. Hydrolysis of the lipid with alkali gave a mixture of fatty acids, glycerol 2-phosphate, sn-glycerol 3-phosphate and O-alpha-d-glucopyranosyl-(1-->2)-O-alpha- d-glucopyranosyl-(1-->1)-d-glycerol. 5. The lipid was unaffected on incubation with phospholipases A, C and D. 6. Diglucosyl diglyceride was isolated after treatment of the lipid with 60% HF, establishing the location of the fatty acid residues. 7. Periodate oxidation studies showed that the sn-glycerol 3-phosphate was esterified to the 6-hydroxyl group of one of the glucose residues in diglucosyl diglyceride.

Acholeplasma laidlawii↗
PubMed1970

Phospholipids of Streptococcus faecalis.

Autoradiograms of total lipid extracts from Streptococcus faecalis ATCC 9790, harvested in the stationary phase from a medium containing (32)P-orthophosphate, showed six major spots. The corresponding compounds were identified as diphosphatidylglycerol (possibly with a penta acyl structure); phosphatidylglycerol; a provisionally identified mixture of alanylphosphatidylglycerol and of the 2'-lysyl-derivative of phosphatidylglycerol; the 3'-lysyl-derivative of phosphatidylglycerol, probably together with some arginylphosphatidylglycerol; a diglucosyl derivative of phosphatidylglycerol; and a compound which was tentatively identified as the 2',3'-dilysyl derivative of phosphatidylglycerol.

Autoradiography↗
PubMedDate not supplied

Crystallization and preliminary X-ray analysis of a lipase from Staphylococcus hyicus.

Single cyrstals of a lipase from Staphylococcus hyicus have been obtained using a combination of 18 to 24% dimethylsulfoxide and 10% isopropanol as a precipitant. The crystals grow at 4 degrees C in 2-3 months. They belong to the orthorhombic space group P212121 with a = 73.31 A, b = 77.96 A, and c = 169.81 A, with two protein molecules per asymmetrical unit. The crystals diffract to at least 2.8 A resolution and are suitable for an X-ray structure analysis.

Crystallization↗
PubMedDate not supplied

Sequence comparison of outer membrane phospholipases A: implications for structure and for the catalytic mechanism.

In this study, the nucleotide sequence of the Enterobacter agglomerans pldA gene encoding outer membrane phospholipase A (OMPLA; EC 3.1.1.32) was determined. Five other OMPLA amino acid sequences have previously been described, and screening of data bases of whole genome sequencing projects revealed the presence of proteins with homology to OMPLA in Helicobacter pylori, Campylobacter jejuni, Yersinia pestis, Neisseria menigitidis and Neisseria gonorrhoeae. Comparison of these eleven OMPLA amino acid sequences revealed that 30 amino acid residues are completely conserved. Implications of the sequence comparison for the catalytic mechanism of OMPLA are discussed. The presence of proteins homologous to OMPLA even in non-enterobacterial Gram-negative bacteria indicates an important physiological role of this enzyme.

Amino Acid Sequence↗