Congenital insulin resistance associated with a conformational alteration in a conserved β-sheet in the insulin receptor L1 domain

被引:19
作者
Rouard, M
Bass, J
Grigorescu, F
Garrett, TPJ
Ward, CW
Lipkind, G
Jaffiole, C
Steiner, DF
Bell, GI
机构
[1] Univ Chicago, Howard Hughes Med Inst, Chicago, IL 60637 USA
[2] Univ Chicago, Dept Med, Chicago, IL 60637 USA
[3] Univ Chicago, Dept Biochem & Mol Biol, Chicago, IL 60637 USA
[4] Inst Univ Rech, Mol Endocrinol Lab, F-34093 Montpellier, France
[5] CSIRO, Div Mol Sci, Parkville, Vic 3052, Australia
[6] Biomol Res Inst, Parkville, Vic 3052, Australia
关键词
D O I
10.1074/jbc.274.26.18487
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
The hormone binding site of members of the insulin receptor family is contained within a highly conserved extracellular region of the receptor. Recent crystallization of the N-terminal region of the binding site revealed two large domains (L1, L2), each organized as a single-stranded right-handed beta-helix, connected by a rod-shaped cysteine-rich domain. Here, we analyze two new naturally occurring mutations in a single p-sheet within L1, D59G and L62P, that we previously identified in a young woman with classic congenital insulin resistance (type A). Substitution of D59G, a beta-sheet connecting loop residue, caused decreased hormone binding but did not disrupt overall folding, assembly, or movement to the cell surface. In contrast, replacement of the adjacent residue L62P, which is located within the beta-sheet, and positioned in a hormone binding surface, completely disrupted intracellular folding, oligomerization, and trafficking and resulted in aberrant proteolytic degradation. Immunohistochemistry in combination with biosynthetic studies showed that misfolded receptors were retained in an incorrect cellular location and that they colocalized with the resident endoplasmic reticulum chaperone calnexin, This study, together with other mutagenesis data, shows that formation of beta-sheet elements within the L1 beta-helix are critical for the folding of the entire extracellular domain of the receptor and that the hormone contact site is composed in part by residues in this domain.
引用
收藏
页码:18487 / 18491
页数:5
相关论文
共 27 条
[1]  
ANDERSEN AS, 1992, J BIOL CHEM, V267, P13681
[2]   ON THE TERTIARY STRUCTURE OF THE EXTRACELLULAR DOMAINS OF THE EPIDERMAL GROWTH-FACTOR AND INSULIN-RECEPTORS [J].
BAJAJ, M ;
WATERFIELD, MD ;
SCHLESSINGER, J ;
TAYLOR, WR ;
BLUNDELL, T .
BIOCHIMICA ET BIOPHYSICA ACTA, 1987, 916 (02) :220-226
[3]   Fusion of insulin receptor ectodomains to immunoglobulin constant domains reproduces high-affinity insulin binding in vitro [J].
Bass, J ;
Kurose, T ;
Pashmforoush, M ;
Steiner, DF .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (32) :19367-19375
[4]   Folding of insulin receptor monomers is facilitated by the molecular chaperones calnexin and calreticulin and impaired by rapid dimerization [J].
Bass, J ;
Chiu, G ;
Argon, Y ;
Steiner, DF .
JOURNAL OF CELL BIOLOGY, 1998, 141 (03) :637-646
[5]  
CREIGHTON TE, 1993, PROTEINS STRUCTURES, P110
[6]   Crystal structure of the first three domains of the type-1 insulin-like growth factor receptor [J].
Garrett, TPJ ;
McKern, NM ;
Lou, MZ ;
Frenkel, MJ ;
Bentley, JD ;
Lovrecz, GO ;
Elleman, TC ;
Cosgrove, LJ ;
Ward, CW .
NATURE, 1998, 394 (6691) :395-399
[7]  
GUSTAFSON TA, 1990, J BIOL CHEM, V265, P18663
[8]  
KJELDSEN T, 1994, J BIOL CHEM, V269, P32942
[9]  
KRAULIS P, 1991, J APP CRYST A, V47, P110
[10]   Expression and characterization of a 70-kDa fragment of the insulin receptor that binds insulin -: Minimizing ligand binding domain of the insulin receptor [J].
Kristensen, C ;
Wiberg, FC ;
Schäffer, L ;
Andersen, AS .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (28) :17780-17786