Mycoplasma arthritidis-derived mitogen (MAM), a bacterial superantigen, has been crystallized in complex with its human receptor, major histocompatibility complex (MHC) class II antigen, by the hanging-drop vapor-diffusion method. Crystals were obtained under three conditions, with ammonium sulfate, phosphate salt and PEG 8000 as the precipitant. The crystals grown under these conditions all belong to space group I222, with the same unit-cell parameters: a = 137.4, b = 178.2, c = 179.6 A. Diffraction data were collected to 3.3 and 3.4 A resolution from crystals of native and selenomethionylated MAM-MHC complexes, respectively. Self- and cross-rotation function calculations suggest the presence of two complex molecules in the asymmetric unit, resulting in a V(M) of 4.0 and a solvent content of 69%. An interpretable electron-density map was produced using a combination of molecular replacement and SAD phasing.

Recombinant human bile salt-stimulated lipase (hBSSL) was expressed in and secreted by Pichia pastoris, an organism exploited for the large-scale production of recombinant (glyco)proteins by bioprocessing technology. The 76.3-kDa glycoprotein was associated with 75-80 Man and a small amount of GlcNAc. hBSSL has one N-glycosylation site at Asn187, which was 38-40% occupied with a Man(10)GlcNAc(2) structure defined previously in Pichia as the oligosaccharide-lipid form of Man(9)GlcNAc(2) trimmed of the middle-arm terminal alpha 1,2-Man and elongated with Man alpha 1,2Man alpha 1,6-disaccharide attached to the lower-arm core alpha 1,3-Man (Trimble et al. [1991], J. Biol. Chem., 266, 22807-22817). The C-terminal 192 residues of hBSSL contain 16 Pro-rich 11-amino-acid repeats, which include 32 Ser/Thr residues as potential O-glycosylation sites. Using hBSSL as a platform to study Pichia's O-glycosylation capabilities, we found that nearly all of these sites were occupied by mannose-containing O-glycans, whose structures, after beta-elimination and purification, were assigned by (1)H NMR and, in some cases, by linkage-specific exoglycosidases and methylation analysis. The most abundant O-glycan was alpha 1,2-mannobiitol (55%), followed by alpha 1,2-mannotriitol (16%) and mannitol (10%) and a lesser amount was alpha 1,2-mannotetraitol. Unexpectedly, Man(5) and Man(6) O-glycans were present, which had the structure Man beta 1,2Man beta 1,2Man alpha 1,2(Man alpha 1,2)(1,2)mannitol. Also a small amount of a phosphorylated Man(6) O-glycan was characterized by MALDI-TOF MS postsource decay analysis as having the reducing-end mannitol disubstituted with a glycosidically linked phosphorylated Man and an unbranched Man(4) polymer elongated from a different mannitol carbon. This is the first report of the synthesis of beta-Man- and phosphate-containing O-linked constituents on glycoproteins synthesized by P. pastoris.

The cDNA microarray is one technological approach that has the potential to accurately measure changes in global mRNA expression levels. We report an assessment of an optimized cDNA microarray platform to generate accurate, precise and reliable data consistent with the objective of using microarrays as an acquisition platform to populate gene expression databases. The study design consisted of two independent evaluations with 70 arrays from two different manufactured lots and used three human tissue sources as samples: placenta, brain and heart. Overall signal response was linear over three orders of magnitude and the sensitivity for any element was estimated to be 2 pg mRNA. The calculated coefficient of variation for differential expression for all non-differentiated elements was 12-14% across the entire signal range and did not vary with array batch or tissue source. The minimum detectable fold change for differential expression was 1.4. Accuracy, in terms of bias (observed minus expected differential expression ratio), was less than 1 part in 10 000 for all non-differentiated elements. The results presented in this report demonstrate the reproducible performance of the cDNA microarray technology platform and the methods provide a useful framework for evaluating other technologies that monitor changes in global mRNA expression.

Annals of the New York Academy of SciencesVolume 732, Issue 1 p. 315-323 Low Molecular Weight Inhibitors in Corneal Ulcerationa RICHARD E. GALARDY, Corresponding Author RICHARD E. GALARDYCorresponding author.Search for more papers by this authorMARIE E. CASSABONNE, MARIE E. CASSABONNE Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorCARLANNE GIESE, CARLANNE GIESE Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorJAMES H. GILBERT, JAMES H. GILBERT Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorFRANCE LAPIERRE, FRANCE LAPIERRE Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorHENRY LOPEZ, HENRY LOPEZ Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorMARY E. SCHAEFER, MARY E. SCHAEFER Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorROBERT STACK, ROBERT STACK Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorMICHAEL SULLIVAN, MICHAEL SULLIVAN Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorBRENT SUMMERS, BRENT SUMMERS Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorROB TRESSLER, ROB TRESSLER Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorDAVE TYRRELL, DAVE TYRRELL Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorJENNIFER WEE, JENNIFER WEE Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorSCOTT D. ALLEN, SCOTT D. ALLEN Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorJOHN J. CASTELLOT, JOHN J. CASTELLOT Department of Anatomy and Cell Biology, Tufts University School of Medicine, Boston, Massachusetts 02111.Search for more papers by this authorJOHN P. BARLETTA, JOHN P. BARLETTA Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorGREGORY S. SCHULTZ, GREGORY S. SCHULTZ University of Florida, Gainesville, Florida 32610.Search for more papers by this authorLEONARDO A. FERNANDEZ, LEONARDO A. FERNANDEZ Department of Diagnostic Radiology, Yale University School of Medicine, New Haven, Connecticut 06510.Search for more papers by this authorSUSAN FISHER, SUSAN FISHER Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorTIAN-YI CUI, TIAN-YI CUI Department of Stomatology, University of California, San Francisco, California 94143.Search for more papers by this authorHARALD G. FOELLMER, HARALD G. FOELLMER Department of Obstetrics and Gynecology, Yale University School of Medicine, New Haven, Connecticut 06437.Search for more papers by this authorDAMIAN GROBELNY, DAMIAN GROBELNY 43 Brassey Avenue, Rosanna 3084, Victoria, Australia.Search for more papers by this authorWALTER M. HOLLERAN, WALTER M. HOLLERAN Department of Dermatology, Veterans Administration Medical Center, 4150 Clement Street, San Francisco, California 94121.Search for more papers by this author RICHARD E. GALARDY, Corresponding Author RICHARD E. GALARDYCorresponding author.Search for more papers by this authorMARIE E. CASSABONNE, MARIE E. CASSABONNE Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorCARLANNE GIESE, CARLANNE GIESE Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorJAMES H. GILBERT, JAMES H. GILBERT Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorFRANCE LAPIERRE, FRANCE LAPIERRE Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorHENRY LOPEZ, HENRY LOPEZ Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorMARY E. SCHAEFER, MARY E. SCHAEFER Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorROBERT STACK, ROBERT STACK Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorMICHAEL SULLIVAN, MICHAEL SULLIVAN Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorBRENT SUMMERS, BRENT SUMMERS Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorROB TRESSLER, ROB TRESSLER Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorDAVE TYRRELL, DAVE TYRRELL Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorJENNIFER WEE, JENNIFER WEE Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorSCOTT D. ALLEN, SCOTT D. ALLEN Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorJOHN J. CASTELLOT, JOHN J. CASTELLOT Department of Anatomy and Cell Biology, Tufts University School of Medicine, Boston, Massachusetts 02111.Search for more papers by this authorJOHN P. BARLETTA, JOHN P. BARLETTA Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorGREGORY S. SCHULTZ, GREGORY S. SCHULTZ University of Florida, Gainesville, Florida 32610.Search for more papers by this authorLEONARDO A. FERNANDEZ, LEONARDO A. FERNANDEZ Department of Diagnostic Radiology, Yale University School of Medicine, New Haven, Connecticut 06510.Search for more papers by this authorSUSAN FISHER, SUSAN FISHER Glycomed Incorporated, 860 Atlantic Avenue, Alameda, California 94501.Search for more papers by this authorTIAN-YI CUI, TIAN-YI CUI Department of Stomatology, University of California, San Francisco, California 94143.Search for more papers by this authorHARALD G. FOELLMER, HARALD G. FOELLMER Department of Obstetrics and Gynecology, Yale University School of Medicine, New Haven, Connecticut 06437.Search for more papers by this authorDAMIAN GROBELNY, DAMIAN GROBELNY 43 Brassey Avenue, Rosanna 3084, Victoria, Australia.Search for more papers by this authorWALTER M. HOLLERAN, WALTER M. HOLLERAN Department of Dermatology, Veterans Administration Medical Center, 4150 Clement Street, San Francisco, California 94121.Search for more papers by this author First published: September 1994 https://doi.org/10.1111/j.1749-6632.1994.tb24746.xCitations: 120 a This work was supported in part by National Institutes of Health grant EY05587. AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Citing Literature Volume732, Issue1Inhibition of Matrix Metalloproteinases: Therapeutic PotentialSeptember 1994Pages 315-323 RelatedInformation