Synthesis of calcineurin-resistant derivatives of FK506 and selection of compensatory receptors

被引:36
作者
Clemons, PA
Gladstone, BG
Seth, A
Chao, ED
Foley, MA
Schreiber, SL
机构
[1] Harvard Univ, Howard Hughes Med Inst, Cambridge, MA 02138 USA
[2] Harvard Univ, Dept Mol & Cellular Biol, Cambridge, MA 02138 USA
[3] Harvard Univ, Dept Chem & Chem Biol, Cambridge, MA 02138 USA
[4] Harvard Univ, Sch Med, Inst Chem & Cell Biol, Boston, MA 02115 USA
来源
CHEMISTRY & BIOLOGY | 2002年 / 9卷 / 01期
关键词
D O I
10.1016/S1074-5521(02)00085-6
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 used olefin metathesis to synthesize C40 derivatives of FK506 and measured their ability, when complexed to FKBP12, to inhibit calcineurin's phosphatase activity. We identified modular dimerization domains (CABS) containing segments of the calcineurin A and B polypeptides. These CABs respond to FK506 both when overexpressed in mammalian cells and in yeast or mammalian three-hybrid assays. Using chemical genetic selection, we identified compensatory mutant CABs that respond to a calcineurin-resistant FK506 derivative at concentrations well below the response threshold for CABS containing only wild-type calcineurin sequence. These reagents provide a small molecule protein combination orthogonal to existing dimerizer systems and may be used with existing systems to increase the complexity of induced-proximity experiments. This new use of the "bump-hole" strategy protects target cells from complications arising from the inhibition of endogenous calcineurin.
引用
收藏
页码:49 / 61
页数:13
相关论文
共 63 条
[41]  
Luban Jeremy, 1995, Current Opinion in Biotechnology, V6, P59, DOI 10.1016/0958-1669(95)80010-7
[42]   Oligomerization activates c-Raf-1 through a Ras-dependent mechanism [J].
Luo, ZJ ;
Tzivion, G ;
Belshaw, PJ ;
Vavvas, D ;
Marshall, M ;
Avruch, J .
NATURE, 1996, 383 (6596) :181-185
[43]   A NEW CLASS OF YEAST TRANSCRIPTIONAL ACTIVATORS [J].
MA, J ;
PTASHNE, M .
CELL, 1987, 51 (01) :113-119
[44]   Calcium triggers calcineurin-dependent synaptic vesicle recycling in mammalian nerve terminals [J].
Marks, B ;
McMahon, HT .
CURRENT BIOLOGY, 1998, 8 (13) :740-749
[45]   PKA phosphorylation dissociates FKBP12.6 from the calcium release channel (ryanodine receptor): Defective regulation in failing hearts [J].
Marx, SO ;
Reiken, S ;
Hisamatsu, Y ;
Jayaraman, T ;
Burkhoff, D ;
Rosemblit, N ;
Marks, AR .
CELL, 2000, 101 (04) :365-376
[46]  
Muthuswamy SK, 1999, MOL CELL BIOL, V19, P6845
[47]  
NORTHROP JP, 1993, J BIOL CHEM, V268, P2917
[48]   NOVEL VECTORS FOR EXPRESSION OF CDNA-ENCODING EPITOPE-TAGGED PROTEINS IN MAMMALIAN-CELLS [J].
PATI, UK .
GENE, 1992, 114 (02) :285-288
[49]   THE EF-HAND FAMILY OF CALCIUM-MODULATED PROTEINS [J].
PERSECHINI, A ;
MONCRIEF, ND ;
KRETSINGER, RH .
TRENDS IN NEUROSCIENCES, 1989, 12 (11) :462-467
[50]  
Pruschy M N, 1994, Chem Biol, V1, P163, DOI 10.1016/1074-5521(94)90006-X