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The seminal plasma comprises secretions from various accessory sex glands. During fertilization spermatozoa undergo complex sequences of precisely timed events that are regulated by the activation of different intracellular signaling pathways. The precision and efficacy of these pathways are often influenced by the assembly and interactions of multiprotein complexes, thereby directing the flow of regulatory information. Our knowledge about these protein complexes present in human seminal plasma (HuSP) is limited. Here we report the identification and characterization of a native high molecular weight zinc‐binding multiprotein complex from HuSP by utilizing 2‐DE followed by MS. Twenty‐six proteins representing isoforms and/or fragments of 11 different proteins were found to be assembled in this complex. Prostate‐specific antigen, zinc α2‐glycoprotein, prostatic acid phosphatase, and prolactin inducible …