We performed large-scale mutational screening of 129 known deafness-related genes and identified three disease-segregating mutations in the TMPRSS3 gene: two previously reported missense mutations, c.916G>A (p.Ala306Thr) and c.316C>T (p.Arg106Cys), and one novel mutation, c.36delC [p.(Phe13Serfs∗12)]. Here, TMPRSS3 is linked to deafness.