Endometriotic lesions synthesize and secrete a haptoglobin-like protein.
To explore the identity and possible function of endometriosis protein-I (ENDO-I), which is an acidic glycoprotein synthesized and secreted by endometriotic lesions, partial amino acid sequence and cDNA sequence were determined. Partially purified, de novo-synthesized rat endometriosis glycoproteins were separated by two-dimensional SDS-PAGE, transferred to polyvinyl difluoride membranes, and stained with Coomassie blue. Protein corresponding to the size and pI of ENDO-I was cut from the membranes and analyzed by automated Edman degradation. ENDO-I amino acid sequence analysis identified 15 residues that shared significant homology with the beta-chain of rat, mouse, and human haptoglobin (Hp) and human Hp-related protein. Western blot analyses using anti-Hp antibody demonstrated cross-reactivity with de novo-synthesized ENDO-I protein in endometriosis culture media. For nucleotide sequence analysis, poly A-enriched mRNA was isolated from rat endometriotic tissues. A gene-specific oligonucleotide primer was designed and used for 3' rapid amplification of cDNA ends (RACE). Automated sequencing of RACE cDNA fragments identified 859 base pairs, of which 858 were identical to rat Hp. Reverse transcription-polymerase chain reaction was used to demonstrate that ENDO-I transcripts are differentially expressed by endometriosis but not by uterine tissues. In the human, distinct subtypes of Hp as well as proteins sharing epitopes with Hp have been used to diagnose a variety of diseases; therefore, Hp-like ENDO-I may prove to be a nonsurgical diagnostic tool to assess endometriosis. Hepatic Hp, induced by acute-phase stimuli, modulates macrophage function and angiogenic activity. If ENDO-I possesses similar activities, it may be involved with anomalies of the immune system or the etiology and pathophysiology of endometriosis.
Sharpe-Timms
K L
KL
Department of Obstetrics and Gynecology, The University of Missouri, Columbia 65212, USA. kathy@ob.missouri.edu
Piva
M
M
Ricke
E A
EA
Surewicz
K
K
Zhang
Y L
YL
Zimmer
R L
RL
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D018076:DNA, Complementary / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D004715:Endometriosis / Q000378:metabolism*; D005260:Female; D015870:Gene Expression; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D006242:Haptoglobins / Q000737:chemistry / Q000235:genetics; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D051381:Rats; D017421:Sequence Analysis; D017385:Sequence Homology
D018076:DNA, Complementary; D006023:Glycoproteins; D006242:Haptoglobins; C112213:endometriosis protein-1