PARTIAL-PURIFICATION AND AMINO-ACID-SEQUENCE ANALYSIS OF ENDOMETRIOSIS PROTEIN-II (ENDO-II) REVEALS HOMOLOGY WITH TISSUE INHIBITOR OF METALLOPROTEINASES-1 (TIMP-1)

De novo synthesized endometriosis protein-Il (ENDO-II; M, 28,000 to 32 ,000; pI 7.0 to 9.0) was partially purified from rat endometriotic tis sue culture media using affinity chromatography and two-dimensional SD S-PAGE. The protein was electrophoretically transferred to polyvinyl d ifluoride membranes which were stained with Coomassie blue R-250. The stained protein corresponding to ENDO-LI (M, 28,000 to 32,000; pi 7.0 to 9.0) was cut from the membranes for amino acid sequencing. Partial amino acid sequence was determined by automated Edman degradation usin g a gas phase sequencer and phenylthiohydantoin analyzer. Sequence ana lysis of ENDO-II yielded 25 residues: C S C A P T H P Q T A F C N S D L V I R A K F M G. Comparison to sequence databanks demonstrated signi ficant homology with rat (100 %) and human (84 %) tissue inhibitor of metalloproteinases-1 (TIMP-1). Western blot analysis using a TIMP-1 an tibody confirmed amino acid sequence analysis. In conclusion, ENDO-II shares sequence homology with TIMP-1 and cross-reactivity with TIMP-1 antibody and subsequently identifies production of TIMP-1 by endometri otic tissues. The synthesis and secretion of TIMP-1 by endometriosis m ay derange the normal proteolytic milieu of the peritoneal cavity and contribute to the etiology and underlying physiological sequelae assoc iated with the disease endometriosis. (Supported by NICHD 29026)