With Ultra DMA onward, position does matter.
I believe the 80-pin conductor is actually made so that only slaves will be in the middle. Something about one of the pins being empty for that connector.
What master/slave does is arbitrate who gets to use the cable. The slave drive will always check to see if the master drive is currently using the cable. If not, then the slave will begin transmitting. With the old pre-UDMA EIDE spec, I think arbitration was more along the lines of ethernet, but I'm pretty sure that nowadays, master/slave actually means something. Plus, the master drive provides termination for the cable, which is why it's really not a good idea to stick a lone drive in the middle of the cable.