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Identification and purification of proteins from germ cell‐conditioned medium (GCCM)
Author(s) -
Chung Sanny S. W.,
Mruk Dolores,
Lee Will M.,
Cheng C. Yan
Publication year - 1999
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216549900201513
Subject(s) - germ cell , polyacrylamide gel electrophoresis , cell , biology , gel electrophoresis , sodium dodecyl sulfate , protein purification , biochemistry , chemistry , gene , enzyme
Germ cells are known to regulate Sertoli cell and testicutar function possibly through released factor(s) or via cell‐cell contact. However, the identities of many of these putative biological factors are not known. The aim of this study is to present a strategy to identify and purify germ cell‐derived proteins found in germ cell‐conditioned medium (GCCM) at a quantity sufficient to permit protein microsequencing. The purification scheme of a novel germ cell‐derived protein from GCCM designated GC‐26 is presented along with several germ cell proteins using a combination of high pressure liquid chromatography (HPLC) columns. The purity of GC‐26 and other germ cell proteins were confirmed by sodium dodecyl sulfate (SDS)‐polyacrylamide gel electrophoresis (PAGE) and silver staining. The identities of GC‐26, a 26‐kDa polypeptide, and other proteins were determined by direct protein microsequencing. These partial NH2‐terminal amino acid sequences were compared with the existing databases at Protein Identification Resource (PIR), GenBank, and BLAST. These analyses revealed that these proteins are unique. This strategy should be useful for the micropurification of proteins from other biological samples and/or fluids.