Identification and partial purification of a chromatin bound calmodulin activated histone 3 kinase from calf thymus.

A calcium-calmodulin (Ca2(+)-CaM) stimulated histone H3 phosphorylating activity was identified as a component of a nuclear protein complex purified from a 150 mM NaCl extract of calf thymus chromatin. This activity bound to a CaM-Sepharose affinity column in a Ca2+ dependent manner and was eluted off the column in the presence of EGTA. Equilibrium centrifugation of the EGTA eluate on a sucrose density gradient revealed that the activity is a component of a larger complex identified at 25% sucrose.

Bile acids: antioxidants or enhancers of peroxidation depending on lipid concentration.

Using three different assay systems, we have discovered a heretofore unrecognized antioxidant property of bile acids at physiological concentrations. Bile acids inhibit peroxidation of the polyunsaturated lipid, linoleic acid, and of the highly fluorescent protein phycoerythrin. In part, the antioxidant activity results from scavenging of peroxyl radicals by direct oxidation of the bile acids.

Phycoerythrin fluorescence-based assay for peroxy radicals: a screen for biologically relevant protective agents.

Under the conditions of this assay, antioxidants that react rapidly with peroxy free radicals (e.g., ascorbate, vitamin E analogs, urate), protect phycoerythrin completely from damage by such radicals generated by thermal decomposition of 2,2'-azobis(2-amidinopropane); other compounds provide partial concentration-dependent protection.

Monoclonal antibody toward lysosomal cathepsin B cross-reacts preferentially with distinct histone classes.

1. A set of monoclonal antibodies (Mab) was prepared against cathepsin B (CB) from rat preputial-gland, an organ characterized by rapidly-renewing cell populations, which is a uniquely enriched source of lysosomal enzymes, including CB. Minute amounts of CB are known to be transferred abruptly to the nuclear compartment in a variety of activated cells.

Determination of estrogen and androgen receptors in Trichomonas vaginalis and the effects of antihormones.

Trichomonas vaginalis, a common genital pathogen, was found to possess both specific estrogen and specific androgen receptors. These 4.3 S macromolecules were proteinaceous in nature.

Accessibility of tyrosyl residues altered by formation of the histone 2A/2B complex.

The availability of tyrosyl residues to surface iodination was analyzed for histone 2A (H2A), histone 2B (H2B), and the H2A/H2B complex. When H2A is free in solution (200 mM NaCl, pH 7.4) tyrosine-39 and one or both tyrosines-50 and -57 were readily iodinated.

Contact site of histones 2A and 2B in chromatin and in solution.

Irradiation of isolated nuclei or of a complex of histones 2A (H2A) and 2B (H2B) with ultraviolet light produces a covalent cross-link between H2A and H2B. Sequence analysis of the peptides isolated from the H2A-H2B dimer formed in solution and in nuclei demonstrated that both dimers are produced through the covalent linkage of Tyr-40 of H2B and Pro-26 of H2A. Tyrosyl residues proximal to Tyr-40 did not produce a cross-link with H2A, thereby indicating that strict conformational parameters are required for production of the H2A-H2B cross-link.

Primary structures of six antimicrobial peptides of rabbit peritoneal neutrophils.

Six microbicidal peptides, NP-1, NP-2, NP-3a, NP-3b, NP-4, and NP-5, from rabbit peritoneal neutrophils were characterized. As a family, the peptides were 32-34 residues in length, were cystine- and arginine-rich, and each contained three intramolecular disulfide bonds. Within their sequences, the six peptides shared 11-residue positions, which included the six half-cystines contained in each peptide.

Isolation, purification, and amino acid composition of the tunicate hemocyte Thy-1 homolog.

A serologic cross-reacting homolog to rodent Thy-1 glycoproteins has been isolated from hemocyte cell surfaces of the advanced invertebrate group of tunicates. The Thy-1.1 cross-reacting antigenic activity was followed during purification by inhibiting the binding of MRC OX7 monoclonal antibody to pure rat brain Thy-1 in a soluble phase radioimmunoassay.

Antiandrogen and antineoplastic effects of a novel drug, CPC10997.

CPC10997 was found to be effective in vitro as an antiandrogen without effects on either the estrogen or the progesterone receptors in carcinomas of the breast, ovary and prostate as well as in malignant melanomas. Using the clonogenic assay of Salmon et al. [Cancer Treat.

Bilin attachment sites in the alpha and beta subunits of B-phycoerythrin. Amino acid sequence studies.

The amino acid sequence about the sites of attachment of all of the bilin prosthetic groups of the alpha and beta subunits of Porphyridium cruentum B-phycoerythrin has been determined. The sequences of five unique tryptic peptides, each carrying one phycoerythrobilin, are as follows: alpha-1 Ile-Asx-Lys-Cys*-Tyr-Arg alpha-2 Asx-Arg-Leu-Cys*-Val-Pro-Arg beta-1 Met-Ala-Ala-Cys*-Leu-Arg beta-2 Met-Ser-Phe-Ala-Ala-Gly-Asp-Cys*-Thr-Ser-Leu-Ala-Ser-Glu-Val-Ala-Gln-Tyr - Phe-Asp-Arg beta-3 Leu-Asp-Ala-Val-Asn-Ser-Ile-Val-Ser-Asn-Ala-Ser-Cys*-Met-Val-Ser-Asp-Ala - Val-Ser-Gly-Met-Ile-Cys*-Glu-Asn-Pro-Gly-Leu-Ile-Ser-Pro-Gly-Gly-Asn-Cys -Tyr- Thr-Asn-Arg where the designations alpha and beta refer to the subunit from which the peptide was derived. Cysteinyl residues involved in bilin attachment are indicated with an asterisk.

Purification of bovine bone morphogenetic protein by hydroxyapatite chromatography.

Bovine bone morphogenetic protein (bBMP) induces differentiation of mesenchymal-type cells into cartilage and bone. bBMP has an apparent Mr of 18,500 +/- 500 and represents less than 0.001% of the wet weight of bone tissue.

Primary structures of MCP-1 and MCP-2, natural peptide antibiotics of rabbit lung macrophages.

The microbicidal peptides, MCP-1 and MCP-2, of rabbit alveolar macrophages were purified by an improved procedure that employed preparative gel electrophoresis and high performance liquid chromatography. The peptides were arginine- and cystine-rich and lacked free sulfhydryl groups and detectable levels of carbohydrate. Complete sequence determinations revealed that MCP-1 differed from MCP-2 only by the substitution of arginine for leucine at residue 13 from the NH2 terminus and that the molecules were each single chain polypeptides of 33 amino acid residues containing three intramolecular disulfide bonds.

Histone-like protein in the Archaebacterium Sulfolobus acidocaldarius.

The Archaebacterium Thermoplasma acidophilum contains a basic chromosomal protein remarkably similar to the histones of eukaryotes. Therefore, it was of interest to examine a different Archaebacterium for similar proteins. We chose to examine Sulfolobus acidocaldarius because it is thermophilic, like T.

Human bone morphogenetic protein (hBMP).

Human bone morphogenetic protein (hBMP) was chemically extracted from demineralized gelatinized cortical bone matrix by means of a CaCl2 X urea inorganic-organic solvent mixture, differential precipitation in guanidine hydrochloride, and preparative gel electrophoresis. hBMP is isolated in quantities of 1 mg/kg of wet weight of fresh bone, and has the amino-acid composition of an acidic polypeptide. The mol wt is 17 to 18 k-Da (kilodaltons).

Estrogen and progesterone receptors in ovarian neoplasms.

The cytoplasmic receptors for 17 beta-estradiol (ER) and progesterone (PR) were measured in 39 malignant and 15 benign ovarian neoplasms. All eight endometroid carcinomas had positive ER sites, one-half contained PR. The number of ER binding sites decreased as tumor grade increased.

Bone cell differentiation and growth factors.

Bone morphogenetic protein and bone-derived growth factors are biochemical tools for research on induced cell differentiation and local mechanisms controlling cell proliferation. Bone morphogenetic protein irreversibly induces differentiation of perivascular mesenchymal-type cells into osteoprogenitor cells. Bone-derived growth factors are secreted by and for osteoprogenitor cells and stimulate DNA synthesis.

Estrogen and progesterone receptor sites in malignancies of the uterine cervix, vagina, and vulva.

Cytoplasmic receptors for 17 beta-estradiol (ER) and progesterone (PR) were measured in uterine cervical, vaginal, and vulvar carcinomas by the dextran-coated charcoal (DCC) technique. Tissues from 30 patients with cervical carcinoma were examined. Thirteen percent (2 of 16) of well-differentiated squamous carcinomas had positive ER, and 19% (3 of 19) had positive PR.

Nuclear sap protein kinases from calf thymus. Substrate inhibition with histones.

1. Protamines were preferentially phosphorylated by the six protein kinase fractions isolated from calf thymus nuclear sap, but histones with the exception of H4 also proved to be acceptable substrates. 2.

A bovine low molecular weight bone morphogenetic protein (BMP) fraction.

A low MW bone morphogenetic protein fraction (BMP) is quantitatively extracted from bovine bone matrix by an inorganic-organic CaCl2-urea solvent mixture and fractionated by ion exchange and gel chromatography. The BMP fraction induces differentiation of perivascular mesenchymal type cells into cartilage and bone inside the mouse's thigh, outside of double walled diffusion chamber, in muscle pouches in the rabbit anterior abdominal wall, and in 0.8 cm trephine defects in the rat's skull.

The amino acid sequence of the beta subunit of allophycocyanin.

The complete amino acid sequence of the beta subunit of Anabaena variabilis allophycocyanin is: H2N-Ala-Gln-Asp-Ala-Ile-Thr-Ala-Val-Ile-Asn-Ser-Ala-Asp-Val-Gln-Gly-Lys-Tyr-Leu-Asp-Thr-Ala-Ala-Leu-Glu-Lys-Leu-Lys-Ala-Tyr-Phe-Ser-Thr-Gly-Glu-Leu-Arg-Val-Arg-Ala-Ala-Thr-Thr-Ile-Ser-Ala-Asn-Ala-Ala-Ala-Ile-Val-Lys-Glu-Ala-Val-Ala-Lys-Ser-Leu-Leu-Tyr-Ser-Asp-Ile-Thr-Arg-Pro-Gly-Gly-Asn-Met-Tyr-Thr-Thr-Arg-Arg-Tyr-Ala-Ala-Cys-Ile-Arg-Asp-Leu-Asp-Tyr-Tyr-Leu-Arg-Tyr-Ala-Thr-Tyr-Ala-Met-Leu-Ala-Gly-Asp-Pro-Ser-Ile-Leu-Asp-Glu-Arg-Val-Leu-Asn-Gly-Leu-Lys-Glu-Thr-Tyr-Asn-Ser-Leu-Gly-Val-Pro-Val-Gly-Ala-Thr-Val-Gln-Ala-Ile-Gln-Ala-Ile-Lys-Glu-Val-Thr-Ala-Ser-Leu-Val-Gly-Ala-Asp-Ala-Gly-Lys-Glu-Met-Gly-Ile-Tyr-Leu-Asp-Tyr-Ile-Ser-Ser-Gly-Leu-Ser-COOH Phycocyanobilin is attached though a thioether linkage to cysteinyl residue 81, indicated by an asterisk. Comparison of this sequence with those of C-phycocyanins shows that there are 60 identities between corresponding subunits of these two biliproteins. Of the region between residues 79 and 120, 29 residues are identical in the beta subunits of allophycocyanin and phycocyanin.

A histone-like protein (HTa) from Thermoplasma acidophilum. I. Purification and properties.

A histone-like protein (HTa) has been isolated from cell extracts of Thermoplasma acidophilum by column chromatography on DNA-cellulose, hydroxylapatite, and Sephadex G-75, HTa elutes from DNA-cellulose in two fractions, one of which contains an 80-residue form of the protein with an NH2-terminal sequence of Val-Gly. The other fraction apparently contains the 89-residue species, in addition to a 90-residue form of the protein with the NH2-terminal sequence Met-Val. The sequence of 47 residues from the NH2 terminus of the 89-residue protein was established by automated Edman degradation.

A histone-like protein (HTa) from Thermoplasma acidophilum. II. Complete amino acid sequence.

The complete amino acid sequence of the DNA-binding histone-like protein (Hta) from Thermoplasma acidophilum has been established by sequence studies directly on the protein and on tryptic, chymotryptic, and thermolysin peptides derived from the protein. The sequence of the 89-residue form of HTa is: H2N-Val-Gly-Ile-Ser-Glu-Leu-Ser-Lys-Glu-Val-Ala-Lys-Lys-Ala-Asn-Thr-Thr-Gln-Lys -Val-Ala-Arg-Thr-Val-Ile-Lys-Ser-Phe-Leu-Asp-Glu-Ile-Val-Ser-Glu-Ala-Asn-Gly-Gl y-Gln-Lys-Ile-Asn-Leu-Ala-Gly-Phe-Gly-Ile-Phe-Glu-Arg-Arg-Thr-Gln-Gly-Pro-Arg-L ys-Ala-Arg-Asn-Pro-Gln-Thr-Lys-Lys-Val-Ile-Glu-Val-Pro-Ser-Lys-Lys-Lys-Phe-Val- Phe-Arg-Ala-Ser-Ser-Lys-Ile-Lys-Tyr-Gln-Gln-COOH The molecular weight calculated from the sequence is 9,934. Another form of HTa probably differs only by the presence of an additional residue (methionine) at the NH2 terminus (the calculated molecular weight of this form is 10,065).

Thermoplasma acidophilum histone-like protein. Partial amino acid sequence suggestive of homology to eukaryotic histones.

Thermoplasma acidophilum is a freeliving mycoplasma-like organism that has a small basic protein tightly bound to its DNA. The N-terminal sequence of this protein has been determined. It has a distanct but statistically significant homology to eukaryotic histones H2A, H3, and to Escherichia coli protein HU.