Secreted proteins of the human pathogen Corynebacterium diphtheriae might be involved in important pathogen–host cell interactions. Here, we present the first systematic reference map of the extracellular and cell surface proteome fractions of the type strain C. diphtheriae C7_s(‐)^tox‐. The analysis window of 2‐DE covered the pI range from 3 to 10 along with a MW range from 8 to 150 kDa. Computational analysis of the 2‐D gels detected almost 150 protein spots in the extracellular proteome fraction and about 80 protein spots of the cell surface proteome. MALDI‐TOF‐MS and PMF with trypsin unambiguously identified 107 extracellular protein spots and 53 protein spots of the cell surface, representing in total 85 different proteins of C. diphtheriae C7_s(‐)^tox‐. Several of the identified proteins are encoded by pathogenicity islands and might represent virulence factors of C. diphtheriae. Additionally, four solute‐binding proteins (HmuT, Irp6A, CiuA, and FrgD) of different iron ABC transporters were identified, with the hitherto uncharacterized FrgD protein being the most abundant one of the cell surface proteome of C. diphtheriae C7_s(‐)^tox‐.