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J E Mole

Publications and source records attributed to J E Mole.

50 records · Page 3Linked to original sources
PubMed1980

Partial amino acid sequence of human factor D:homology with serine proteases.

Human factor D purified to homogeneity by a modified procedure was subjected to NH2-terminal amino acid sequence analysis by using a modified automated Beckman sequencer. We identified 48 of the first 57 NH2-terminal amino acids in a single sequencer run, using microgram quantities of factor D. The deduced amino acid sequence represents approximately 25% of the primary structure of factor D. This extended NH2-terminal amino acid sequence of factor D was compared to that of other trypsin-related serine proteases. By visual inspection, strong homologies (33--50% identity) were observed with all the serine proteases included in the comparison. Interestingly, factor D showed a higher degree of homology to serine proteases of pancreatic origin than to those of serum origin.

Amino Acid Sequence↗
PubMed1980

Monoclonal and allospecific H-2Kk antisera precipitate a high molecular weight protein.

A protein with an apparent m.w. of 68,000 has been observed in immunoprecipitations of NP-40 solubilized BW5147 thymoma cells by using monoclonal or polyclonal allospecific H-2Kk antisera. By contrast, H-2Dk alloantisera precipitated a 45,000 m.w. protein from the same membrane preparations. The 68,000 m.w. protein is expressed on the plasma membrane as determined by vectoral labeling. Extensive clearing experiments and the use of monoclonal antibody demonstrate that the protein does not bear group-specific C-type viral determinants.

Animals↗
PubMed1978

Purification and characterization of GM1 ganglioside beta-galactosidase from normal feline liver and brain.

GM1 ganglioside beta-galactosidase (GM1-beta-galactosidase) was purified from normal cat brain and liver by a combination of classical and affinity procedures. The final preparation of brain GM1-beta-galactosidase was enriched over 2000-fold with a 36% yield. However, the product was shown to contain several components by disc gel electrophoresis. GM1-beta-galactosidase was also purified from liver with greater than a 30 000-fold enrichment and 40% yield. The liver enzyme was judged homogeneous by disc gel electrophoresis at pH 4.3, 8.1, and 8.9 and by gel chromatography. Both liver and brain GM1-beta-galactosidase(s) eluted as sharp symmetrical peaks from Sephadex G-200 with molecular weights of 250 000 +/- 50 000. The apparent Km determined for 4-methylumbelliferyl beta-D-galactopyranoside (4-MU-Gal) using partially purified brain GM1-beta-galactosidase was 1.73 X 10(-4) M. Liver GM1-beta-galactosidase gave a Km with 4-MU-Gal of 3.25 X 10(-4) M and for [3H]GM1 ganglioside a Km of 4.51 X 10(-4) M was calculated. The pH optima of brain and liver GM1-beta-galactosidase using 4-MU-Gal was 3.8-4.5. By contrast, liver GM1-beta-galactosidase gave a sharp activity peak at pH 4.2 with [3H]GM1 ganglioside. Inhibition by mercuric chloride and sensitivity to hydrogen peroxide and persulfate suggest the involvement of a sulfhydryl in catalysis.

Animals↗
PubMed1977

Primary structure of human J chain: isolation and characterization of tryptic and chymotryptic peptides of human J chain.

Human J chain isolated from the plasma of a patient with Waldenstrom's macroglobulinemia was subjected to extended and limited digestion with trypsin and chymotrypsin. The digests were fractionated by combination of column chromatography and high voltage paper electrophoresis. Peptide purity was established by their amino acid analysis and a single amino terminal residue. All the necessary peptides which would provide the total primary structure of molecule were thus obtained.

Amino Acids↗
PubMed1977

Primary structure of human J chain: alignment of peptides from chemical and enzymatic hydrolyses.

The primary structure of the J chain from a human Waldenströms IgM protein has been determined using a combination of automated and conventional Edman degradative procedures. Eighty-five percent of the sequence was established with peptides isolated from tryptic digests of carboxyamidomethylated and citraconylated J chain, many of which were sequenced completely. Alignment of the tryptic fragments was achieved with peptides generated by chymotrypsin and limited acid hydrolyses. The j chain consits of 129 amino acids and a single oligosaccharide structure linked to asparagine at positon 43 of the sequence. The molecular weight, including 7.5% carbohydrate by weight, is 16 422. The location and arrangement of three half-cystines could be deduced from previous studies, whereas the pairing of the remaining five disulfide bonds still needs to be clarified.

Amino Acid Sequence↗
PubMed1977

Structural analysis of the peptides derived from specific acid-catalyzed hydrolysis at aspartylprolyl peptide bonds in human J chain.

Human J chain from IgM has been selectively cleaved at three aspartylprolyl peptide bonds to yield four fragments containing 62, 20, 25, and 22 amino acids, respectively. The amino acid sequence of each peptide has been partially determined, (59 of a total of 129 residues) and its position in the J chain ascertained. There were no obvious similarities to known sequences in other immunoglobulin polypeptide chains.

Amino Acid Sequence↗
PubMed1975

Evidence for the absence of noncovalent bonds in the Fcmu region of IgM.

IgM isolated from the sera of five patients with Waldenström's macroglobulinemia was subjected to tryptic digestion at 60 degrees C. The Fc5mu fragments recovered from the digests were reduced by 0.05 M cysteine and alkylated by iodoacetamide, producing large quantities of an Fcmu fragment having a sedimentation velocity (see article) of 2.9S and a molecular weight of 33,500 by sedimentation equilibrium in neutral buffer. Further studies on the Fcmu fragment from one of the proteins demonstrated that it was not dissociated into smaller fragments by 5 M guanidine-HC1, even after reduction with 0.1 M 2-mercaptoethanol in 5 M guanidine at pH 7.5. The number of sulfhydryl groups released by the latter treatment indicated the presence of two intrachain disulfide bonds. These observations provide evidence that this portion of the mu-chain demonstrates minimal noncovalent interactions. Tryptic digestion of the Fcmu fragment at 37 degrees C resulted in the production of several lower molecular weight fractions. The three major fractions demonstrated apparent molecular weights of 21,000, 13,800 and 6800 by sedimentation equilibrium in 5 M guanidine-HC1. The latter fraction (fraction C) had no detectable carbohydrate and consisted of two disulfide-bonded peptides having molecular weights of approximately 3800 and 2200. Studies on the amino acid composition and amino-terminal sequences indicated that fraction C was derived from the Cmu4 homology region and consisted of residues 468 through 546 of the mu-chain with the tryptic peptides encompassing residues 492 through 514 missing.

Amino Acid Sequence↗
PubMedDate not supplied

Plasma and cytoplasmic gelsolins are encoded by a single gene and contain a duplicated actin-binding domain.

Gelsolin is representative of a class of actin-modulating proteins found in lower eukaryotes to mammals, which sever actin filaments. Gelsolin found in the cytoplasm of cells is functionally similar to a mammalian plasma protein of similar size, originally called ADF or brevin. Human plasma and rabbit macrophage gelsolins differ by the presence of a 25-amino-acid residue extension on plasma gelsolin which appears to account for the difference in relative molecular mass (Mr) between the proteins as assessed by SDS-polyacrylamide gel electrophoresis (PAGE), 93,000 (93K) and 90K, respectively. Here we report the isolation of full-length human plasma gelsolin complementary DNA clones from a HepG2 library. The inferred amino-acid sequence reveals the presence of a signal peptide, a long tandem repeat that matches the actin-binding domains of gelsolin, a tetrapeptide present in actin and extended regions of identical sequence with rabbit macrophage gelsolin. Southern blot analysis indicates that a single gene in the haploid genome encodes both protein forms.

Amino Acid Sequence↗