'Human angiotensin-converting enzyme-related carboxypeptidase (ACE2) is a zinc metalloprotease whose closest homolog is angiotensin I-converting enzyme. To begin to elucidate the physiological role of ACE2, ACE2 was purified, and its catalytic activity was characterized. ACE2 proteolytic activity has a pH optimum of 6.5 and is enhanced by monovalent anions, which ... More
hsp90 is required for heme binding and activation of apo-neuronal nitric-oxide synthase: geldanamycin-mediated oxidant generation is unrelated to any action of hsp90.
Authors Billecke Scott S; Bender Andrew T; Kanelakis Kimon C; Murphy Patrick J M; Lowe Ezra R; Kamada Yasuhiko; Pratt William B; Osawa Yoichi;
JournalJ Biol Chem
PubMed ID11923316
'It is established that neuronal NO synthase (nNOS) is associated with the chaperone hsp90, although the functional role for this interaction has not been defined. We have discovered that inhibition of hsp90 by radicicol or geldanamycin nearly prevents the heme-mediated activation and assembly of heme-deficient apo-nNOS in insect cells. This ... More
The major conformational IgE-binding epitopes of hevein (Hev b6.02) are identified by a novel chimera-based allergen epitope mapping strategy.
Authors Karisola Piia; Alenius Harri; Mikkola Jari; Kalkkinen Nisse; Helin Jari; Pentikäinen Olli T; Repo Susanna; Reunala Timo; Turjanmaa Kristiina; Johnson Mark S; Palosuo Timo; Kulomaa Markku S;
JournalJ Biol Chem
PubMed ID11909866
'A novel approach to localize and reconstruct conformational IgE-binding epitope regions of hevein (Hev b6.02), a major natural rubber latex allergen, is described. An antimicrobial protein (AMP) from the amaranth Amaranthus caudatus was used as an immunologically non-IgE-binding adaptor molecule to which terminal or central parts of hevein were fused. ... More
Enhancing yield of infectious Bursal disease virus structural proteins in baculovirus expression systems: focus on media, protease inhibitors, and dissolved oxygen.
Authors Hu Y C; Bentley W E;
JournalBiotechnol Prog
PubMed ID10585191
'Structural proteins of the poultry pathogen, infectious bursal disease virus (IBDV), were expressed in the baculovirus/insect cell expression system. To date, several reports have indicated that animal virus structural proteins are expressed only at low yield in this system. In this article, several factors were examined to enhance yield. These ... More
Crystal Structure of Imaginal Disc Growth Factor-2. A MEMBER OF A NEW FAMILY OF GROWTH-PROMOTING GLYCOPROTEINS FROM DROSOPHILA MELANOGASTER.
Authors Varela Paloma F; Llera Andrea S; Mariuzza Roy A; Tormo Jose;
JournalJ Biol Chem
PubMed ID11821393
'Imaginal disc growth factor-2 (IDGF-2) is a member of a recently described family of Drosophila melanogaster-soluble polypeptide growth factors that promote cell proliferation in imaginal discs. Although their precise mode of action has not been established, IDGFs cooperate with insulin in stimulating the growth of imaginal disc cells. We report ... More
Titration of non-occluded baculovirus using a cell viability assay.
AuthorsMena JA, RamÃrez OT, Palomares LA,
JournalBiotechniques
PubMed ID12613247
Heparin inhibits the binding of beta 2-glycoprotein I to phospholipids and promotes the plasmin-mediated inactivation of this blood protein. Elucidation of the consequences of the two biological events in patients with the anti-phospholipid syndrome.
Authors Guerin Jan; Sheng Yonghua; Reddel Stephen; Iverson G Michael; Chapman Michael G; Krilis Steven A;
JournalJ Biol Chem
PubMed ID11711550
The phospholipid-binding plasma protein beta2-glycoprotein I (beta2-GPI) is the primary antigen recognized by the circulating autoantibodies in patients with the
Mutational analysis of the DEAD-box RNA helicase eIF4AII characterizes its interaction with transformation suppressor Pdcd4 and eIF4GI.
AuthorsZakowicz H, Yang HS, Stark C, Wlodawer A, Laronde-Leblanc N, Colburn NH,
JournalRNA
PubMed ID15661843
Eukaryotic initiation factor (eIF) 4A unwinds secondary and tertiary structures in the 5'-untranslated region of mRNA, permitting translation initiation. Programmed cell death 4 (Pdcd4) is a novel transformation suppressor and eIF4A-binding partner that inhibits eIF4A helicase activity and translation. To elucidate the regions of eIF4A that are functionally significant in ... More
The significance of tetramerization in promoter recruitment by Stat5.
AuthorsJohn S, Vinkemeier U, Soldaini E, Darnell JE Jr, Leonard WJ
JournalMol Cell Biol
PubMed ID10022878
Stat5a and Stat5b are rapidly activated by a wide range of cytokines and growth factors, including interleukin-2 (IL-2). We have previously shown that these signal transducers and activators of transcription (STAT proteins) are key regulatory proteins that bind to two tandem gamma interferon-activated site (GAS) motifs within an IL-2 response ... More
PRODUCTION OF MOSQUITO DENSONUCLEOSIS VIRUSES BY AEDES ALBOPICTUS C6/36 CELLS ADAPTED TO SUSPENSION CULTURE IN SERUM-FREE PROTEIN-FREE MEDIA.
AuthorsSuchman E, Carlson J,
JournalIn Vitro Cell Dev Biol Anim
PubMed ID15311964
Mosquito densonucleosis viruses (MDVs) have the potential for use as biocontrol agents. To facilitate densovirus production, the Aedes albopictus mosquito cell line C6/36 was adapted to two commercially available serum-free protein-free media (SFPFM), Sf-900 II and Drosophila-SFM. Cells adapted more slowly to growth in Sf-900 II medium, but once adapted, ... More
Enhanced growth of Sf-9 cells to a maximum density of 5.2 x 10(7) cells per mL and production of beta-galactosidase at high cell density by fed batch culture.
A novel 43-kDa protein as a negative regulatory component of phenoloxidase-induced melanin synthesis.
AuthorsZhao M, Söderhäll I, Park JW, Ma YG, Osaki T, Ha NC, Wu CF, Söderhäll K, Lee BL,
JournalJ Biol Chem
PubMed ID15857824
The melanization reaction induced by activated phenoloxidase in arthropods is important in the multiple host defense innate immune reactions, leading to the sequestration and killing of invading microorganisms. This reaction ought to be tightly controlled because excessive formation of quinones and systemic hypermelanization are deleterious to the hosts, suggesting that ... More
Comparison of filamin A-induced cross-linking and Arp2/3 complex-mediated branching on the mechanics of actin filaments.
Authors Nakamura Fumihiko; Osborn Eric; Janmey Paul A; Stossel Thomas P;
JournalJ Biol Chem
PubMed ID11786548
We compared the effects of human filamin A (FLNa) and the activated human Arp2/3 complex on mechanical properties of actin filaments. As little as 1 FLNa to 800 polymerizing actin monomers induces a sharp concentration-dependent increase in the apparent viscosity of 24 microm actin, a parameter classically defined as a ... More
Combinatorial expression of GATA4, Nkx2-5, and serum response factor directs early cardiac gene activity.
Herein, the restricted expression of serum response factors (SRF) closely overlapped with Nkx2-5 and GATA4 transcripts in early chick embryos coinciding with the earliest appearance of cardiac alpha-actin (alphaCA) transcripts and nascent myocardial cells. The combinatorial expression of SRF, a MADS box factor Nkx2-5 (a NK4 homeodomain), and/or GATA4, a ... More
Characterization of three fragments that constitute the monomers of the human lysyl hydroxylase isoenzymes 1-3. The 30-kDa N-terminal fragment is not required for lysyl hydroxylase activity.
Lysyl hydroxylase (LH) catalyzes the formation of hydroxylysine in collagens; three human isoenzymes have been cloned so far. We report here on the purification of all three recombinant isoenzymes to homogeneity from the medium of cultured insect cells, and we demonstrate that they are all homodimers. Limited proteolysis experiments identified ... More
High-level expression and purification of secreted forms of herpes simplex virus type 1 glycoprotein gD synthesized by baculovirus-infected insect cells.
Authors Sisk W P; Bradley J D; Leipold R J; Stoltzfus A M; Ponce de Leon M; Hilf M; Peng C; Cohen G H; Eisenberg R J;
JournalJ Virol
PubMed ID8289380
Two forms of herpes simplex virus glycoprotein gD were recombined into Autographa californica nuclear polyhedrosis virus (baculovirus) and expressed in infected Spodoptera frugiperda (Sf9) cells. Each protein was truncated at residue 306 of mature gD. One form, gD-1(306t), contains the coding sequence of Patton strain herpes simplex virus type 1 ... More