The electronic characteristics of organic optoelectronic materials determine the performance of corresponding devices. Clarifying the relationship between molecular structure and electronic characteristics at the single-molecule level can help to achieve high performance for organic optoelectronic materials and devices, especially for organic photovoltaics. In this work, a typical acceptor-donor-acceptor (A-D-A)-type molecule is explored by combining theoretical and experimental studies to reveal the intrinsic electronic characteristics at the single-molecule level. Specifically, the A-D-A-type molecule with 1,1-dicyano methylene-3-indanone (INCN) acceptor units exhibits an enhanced conductance in single-molecule junctions when compared with the control donor molecule, because the acceptor units of the A-D-A-type molecule contribute additional transport channels. In addition, through opening the S center dot center dot center dot O noncovalent conformational lock by protonation to expose the -S anchoring sites, the charge transport of the D central part is detected, proving that the conductive orbitals contributed by the INCN acceptor groups can penetrate the whole A-D-A molecule. These results provide important insights into the development of high-performance organic optoelectronic materials and devices toward practical applications.