Glycosylation affects both the three-dimensional structure and antibody binding properties of the HIV-1(IIIB) GP120 peptide RP135

被引:90
作者
Huang, XL
Barchi, JJ
Lung, EDT
Roller, PP
Nara, PL
Muschik, J
Garrity, RR
机构
[1] NCI, MED CHEM LAB, DIV BASIC SCI, BETHESDA, MD 20892 USA
[2] NCI, VACCINE RESISTANT DIS SECT, DIV BASIC SCI, FREDERICK CANC RES & DEV CTR, FREDERICK, MD 21702 USA
关键词
D O I
10.1021/bi9703655
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We have prepared glycosylated analogues of the principal neutralizing determinant of gp120 and studied their conformations by NMR and circular dichroism spectroscopies. The 24-residue peptide from the HIV-1(IIIB) isolate (residues 308-331) designated RP135, which contains the immunodominant tip of the V3 loop, was glycosylated with both N- and O-linked sugars. The structures of two glycopeptides, one with an N-linked beta-glucosamine (RP135(NG)) and. the other with two O-linked alpha-galactosamine units (RP135(digal)), were studied by NMR and circular dichroism spectroscopies. Molecular dynamics calculations based on the NMR data obtained in water solutions were performed to explore the conformational substates sampled by the glycopeptides. The data showed that covalently linking a carbohydrate to the peptide has a major effect on the local conformation and imparts additional minor changes at more distant sites of partially defined secondary structure. In particular, the transient beta-type turn comprised of the -Gly-Pro-Gly-Arg- segment at the ''tip'' of the V3 loop is more highly populated in RP135(digal) than in the native peptide and N-linked analogue. Binding data for the glycopeptides with 0.5 beta, a monoclonal antibody mapped to the RP135 sequence, revealed a significant enhancement in binding for RP135(digal) as compared with the native peptide, whereas binding was reduced for the N-linked glycopeptide. These data show that glycosylation of V3 loop peptides can affect their conformations as well as their interactions with antibodies, The design of more ordered and biologically relevant conformations of immunogenic regions from gp120 may aid in the design of more effective immunogens for HIV-1 vaccine development.
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页码:10846 / 10856
页数:11
相关论文
共 72 条
[1]   EFFECTS OF GLYCOSYLATION ON PEPTIDE BACKBONE CONFORMATION [J].
ANDREOTTI, AH ;
KAHNE, D .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1993, 115 (08) :3352-3353
[2]  
[Anonymous], THEOR CULT SOC
[3]   MLEV-17-BASED TWO-DIMENSIONAL HOMONUCLEAR MAGNETIZATION TRANSFER SPECTROSCOPY [J].
BAX, A ;
DAVIS, DG .
JOURNAL OF MAGNETIC RESONANCE, 1985, 65 (02) :355-360
[4]   HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 ENVELOPE GLYCOPROTEIN IS MODIFIED BY O-LINKED OLIGOSACCHARIDES [J].
BERNSTEIN, HB ;
TUCKER, SP ;
HUNTER, E ;
SCHUTZBACH, JS ;
COMPANS, RW .
JOURNAL OF VIROLOGY, 1994, 68 (01) :463-468
[5]   CHARMM - A PROGRAM FOR MACROMOLECULAR ENERGY, MINIMIZATION, AND DYNAMICS CALCULATIONS [J].
BROOKS, BR ;
BRUCCOLERI, RE ;
OLAFSON, BD ;
STATES, DJ ;
SWAMINATHAN, S ;
KARPLUS, M .
JOURNAL OF COMPUTATIONAL CHEMISTRY, 1983, 4 (02) :187-217
[6]  
BRUENGER AT, 1993, X PLOR VERSION 3 1 S
[7]   LOCAL AND GLOBAL STRUCTURAL-PROPERTIES OF THE HIV-MN V3 LOOP [J].
CATASTI, P ;
FONTENOT, JD ;
BRADBURY, E ;
GUPTA, G .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (05) :2224-2232
[8]   Structure and polymorphism of HIV-1 third variable loops [J].
Catasti, P ;
Bradbury, EM ;
Gupta, G .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (14) :8236-8242
[9]   SOLUTION CONFORMATIONAL PREFERENCES OF IMMUNOGENIC PEPTIDES DERIVED FROM THE PRINCIPAL NEUTRALIZING DETERMINANT OF THE HIV-1 ENVELOPE GLYCOPROTEIN GP120 [J].
CHANDRASEKHAR, K ;
PROFY, AT ;
DYSON, HJ .
BIOCHEMISTRY, 1991, 30 (38) :9187-9194
[10]   THE V3 LOOPS OF THE HIV-1 AND HIV-2 SURFACE GLYCOPROTEINS CONTAIN PROTEOLYTIC CLEAVAGE SITES - A POSSIBLE FUNCTION IN VIRAL FUSION [J].
CLEMENTS, GJ ;
PRICEJONES, MJ ;
STEPHENS, PE ;
SUTTON, C ;
SCHULZ, TF ;
CLAPHAM, PR ;
MCKEATING, JA ;
MCCLURE, MO ;
THOMSON, S ;
MARSH, M ;
KAY, J ;
WEISS, RA ;
MOORE, JP .
AIDS RESEARCH AND HUMAN RETROVIRUSES, 1991, 7 (01) :3-16