Characterization of mammalian stanniocalcin receptors - Mitochondrial targeting of ligand and receptor for regulation of cellular metabolism

被引:109
作者
McCudden, CR
James, KA
Hasilo, C
Wagner, GF [1 ]
机构
[1] Univ Western Ontario, Fac Med & Dent, Dept Physiol, London, ON N6A 5C1, Canada
[2] Univ Western Ontario, Fac Sci, Dept Biol, London, ON N6A 5C1, Canada
关键词
D O I
10.1074/jbc.M205954200
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 polypeptide hormone stanniocalcin (STC) is widely expressed in mammalian tissues. STC acts locally in kidney and gut to modulate calcium and phosphate excretion, and its overexpression in mice results in high serum phosphate, dwarfism, and increased metabolic rate. STC has also been linked to cancer, pregnancy, lactation, angiogenesis, organogenesis, cerebral ischemia, and hypertonic stress. In this report we have characterized the STC receptor and the functional targeting of ligand and receptor to mitochondria. For receptor binding analysis, a stanniocalcin-alkaline phosphatase fusion protein was engineered. Subsequent binding assays using the fusion protein indicated that kidney and liver contained the highest number of binding sites with affinities of 0.8 and 0.25 nm, respectively. Intriguingly, purified mitochondria from both tissues yielded similar high affinity binding sites. Fractionation analysis revealed that the majority of binding sites were localized to the inner mitochondrial membrane. In further studies, we characterized the time course of STC-alkaline phosphatase fusion protein sequestration by intact mitochondria. In situ ligand binding also revealed discrete, displaceable, binding to plasma membranes and mitochondria of nephron cells and liver hepatocytes. The existence of mitochondrial receptors prompted a similar search for the ligand. Immunogold electron microscopy revealed that STC was preferentially concentrated in the mitochondria of all nephron segments targeted by STC. Subcellular fractionation revealed that >90% of cellular STC immunoreactivity was mitochondrial, confined to the inner matrix, and similar in size to recombinant STC (50 kDa). In functional studies, recombinant STC had concentration-dependent stimulatory effects on electron transfer by sub-mitochondrial particles. Collectively the evidence implies a role for STC in cell metabolism.
引用
收藏
页码:45249 / 45258
页数:10
相关论文
共 48 条
[1]   Stanniocalcin from an ancient teleost: a monomeric form of the hormone and a possible extracorpuscular distribution [J].
Amemiya, Y ;
Marra, LE ;
Reyhani, N ;
Youson, JH .
MOLECULAR AND CELLULAR ENDOCRINOLOGY, 2002, 188 (1-2) :141-150
[2]  
BERGERON JJM, 1978, J BIOL CHEM, V253, P4058
[3]   Analysis of the pathways of nitric oxide utilization in mitochondria [J].
Cadenas, E ;
Poderoso, JJ ;
Antunes, F ;
Boveris, A .
FREE RADICAL RESEARCH, 2000, 33 (06) :747-756
[4]   A NOVEL HUMAN CDNA HIGHLY HOMOLOGOUS TO THE FISH HORMONE STANNIOCALCIN [J].
CHANG, ACM ;
JANOSI, J ;
HULSBEEK, M ;
DEJONG, D ;
JEFFREY, KJ ;
NOBLE, JR ;
REDDEL, RR .
MOLECULAR AND CELLULAR ENDOCRINOLOGY, 1995, 112 (02) :241-247
[5]   Molecular cloning and characterization of mouse stanniocalcin cDNA [J].
Chang, ACM ;
Dunham, MA ;
Jeffrey, KJ ;
Reddel, RR .
MOLECULAR AND CELLULAR ENDOCRINOLOGY, 1996, 124 (1-2) :185-187
[6]   Requirement for transforming growth factor β1 in controlling T cell apoptosis [J].
Chen, WJ ;
Jin, WW ;
Tian, HS ;
Sicurello, P ;
Frank, M ;
Orenstein, JM ;
Wahl, SM .
JOURNAL OF EXPERIMENTAL MEDICINE, 2001, 194 (04) :439-453
[7]   IDENTIFICATION AND CLONING OF ELF-1, A DEVELOPMENTALLY EXPRESSED LIGAND FOR THE MEK4 AND SEK RECEPTOR TYROSINE KINASES [J].
CHENG, HJ ;
FLANAGAN, JG .
CELL, 1994, 79 (01) :157-168
[8]   An alternative transcript of the rat renin gene can result in a truncated prorenin that is transported into adrenal mitochondria [J].
Clausmeyer, S ;
Stürzebecher, R ;
Peters, J .
CIRCULATION RESEARCH, 1999, 84 (03) :337-344
[9]   Development of a human stanniocalcin radioimmunoassay: serum and tissue hormone levels and pharmacokinetics in the rat [J].
De Niu, P ;
Radman, DP ;
Jaworski, EM ;
Deol, H ;
Gentz, R ;
Su, J ;
Olsen, HS ;
Wagner, GF .
MOLECULAR AND CELLULAR ENDOCRINOLOGY, 2000, 162 (1-2) :131-144
[10]   Immunolocalization of stanniocalcin in human kidney [J].
De Niu, P ;
Olsen, HS ;
Gentz, R ;
Wagner, GF .
MOLECULAR AND CELLULAR ENDOCRINOLOGY, 1998, 137 (02) :155-159