Publications (6)

  • Source
    M Gnanasekar · F G Suleman · K Ramaswamy · J D Caldwell
    [Show abstract] [Hide abstract] ABSTRACT: The present study reports the identification of human sex hormone binding globulin (SHBG)-interacting proteins in the brain using a phage display-based screening technology. Phage display is a system in which a foreign protein is displayed on the surface of a bacteriophage as a fusion protein with one of the coat proteins of the bacteriophage. T7 phage clones expressing normal human brain proteins (human normal brain phage-display cDNA expression library) were screened using SHBG as bait. The bound phage clones were then identified by DNA sequencing and by BLAST search analysis. Of the twenty binding proteins analyzed, three were found to be membrane-associated proteins: synaptosomal associated protein 25 (SNAP25), Thy-1 cell surface antigen and zonadhesin. Further studies will determine if the interactions of SHBG with these proteins have any role in the internalization and cell signaling events or whether they contribute to steroid delivery to specific cells.
    Full-text available · Article · Nov 2009 · International Journal of Molecular Medicine
  • J D Caldwell · F Suleman · G F Jirikowski
    Article · Sep 2007 · Hormone and Metabolic Research
  • Source
    [Show abstract] [Hide abstract] ABSTRACT: Sex hormone-binding globulin (SHBG) is a 94-kDa homodimer that binds steroids and is made in the hypothalamus. We have demonstrated that infusions of SHBG into the hypothalami of rats increase their female sexual receptivity except when SHBG is coupled to dihydrotestosterone (DHT) suggesting that SHBG has an active function in behavioral neuroendocrinology. This study examines the possibility that SHBG is internalized by neuronal and/or non-neuronal brain cells as one possible mode of action using in vitro and in vivo techniques. First, analysis of the uptake of radiolabeled SHBG ((125)I-SHBG) found (125)I-SHBG uptake in HT22 hippocampal cells stably transfected with cDNA for ER beta (HT22-ER beta). The addition of DHT to (125)I-SHBG significantly inhibited (125)I-SHBG uptake in HT22-ER beta cells but not in HT22-ER alpha or HT22 wild-type cells. SHBG internalization was specific as it did not occur in either the human neuroblastoma cell line SK-N-SH or the glioma cell line C6. Second, SHBG was labeled with a fluor (Alexa-555), and infused into the lateral cerebroventricles of ovariectomized rats. Optimal SHBG uptake was seen 10 min after these infusions. SHBG uptake was seen in specific parts of the choroid plexus and periventricular cells as well as into cells in the paraventricular nucleus, the medial forebrain bundle, and the habenula. These studies suggest that SHBG is internalized by brain cells, which may be affected by the presence of ER beta. The gonadal steroids have numerous effects in brain and the discovery that the steroid-binding protein SHBG is taken up into neurons and brain cells may demand a change in thinking about how steroids are delivered to brain cells to affect neurophysiology.
    Full-text available · Article · Feb 2007 · Neuroendocrinology
  • J D Caldwell · F Suleman · S H-H Chou · [...] · G F Jirikowski
    Article · May 2006 · Hormone and Metabolic Research
  • Venkata M Reddy · Funmilayo G Suleman
    [Show abstract] [Hide abstract] ABSTRACT: Mycobacterium avium complex (MAC) adheres, invades and multiplies inside epithelial cells. Earlier, we demonstrated two MAC protein adhesins, 25 and 31 kDa, binding with HEp-2 cells. The 25 kDa MAC adhesin was found to be superoxide dismutase (SOD). In this study, epithelial cell (HEp-2 and A549) ligands for MAC-SOD were identified by probing two-dimensional western blots of epithelial extracts with MAC proteins followed by monoclonal anti-MAC-SOD antibodies. Three epithelial cell proteins with molecular masses 43, 40 and 18 kDa, present in both membrane and cytosolic fractions, were found to bind with MAC-SOD. Based on the N-terminal amino acid sequences, the 43, 40 and 18 kDa epithelial proteins were identified as aldolase, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and cyclophilin A (CypA), respectively. Furthermore, MAC-SOD was found to bind to purified rabbit muscle aldolase, GAPDH and recombinant CypA in western blotting.
    Article · Mar 2004 · Microbial Pathogenesis
  • V.M. Reddy · F.G. Suleman · D.A. Hayworth
    [Show abstract] [Hide abstract] ABSTRACT: Mycobacterium avium complex (MAC) is known to colonize the gastrointestinal tract of human immunodeficiency virus (HIV) infected patients before causing bacteremia and disseminated disease. However, the mechanism involved in the gastrointestinal colonization is not known. To identify putative intestinal mucus receptors which serve as anchor for MAC colonization. C57BL/6 mouse intestinal mucus was subjected to single and two-dimensional electrophoresis and blotted on nitrocellulose membranes. MAC specific mucus proteins were identified by probing the mucus western blots with biotinylated proteins derived from M.avium strain 101 (MAC101). Biotinylated MAC 101 proteins recognized a 39 kDa intestinal mucus glycoprotein. The protein displaying an isoelectric point (pI) of 9.0, was found to be periodate sensitive but resistant to sialidase, heparinase I and chondroitinase ABC. The internal amino acid sequence of the 39 kDa protein displayed homology with fructose-1-6-bisphosphate aldolase B (aldolase). The proclivity between MAC adhesins and aldolase was confirmed by probing rabbit muscle aldolase with MAC proteins. Furthermore, both 25 and 31 kDa MAC adhesins, superoxide dismutase and heparin binding protein, respectively, were found to bind to aldolase. MAC binds to intestinal mucus aldolase, conceivably facilitating intestinal colonization of the organism.
    Article · Feb 2004 · Tuberculosis